Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope

Transcription

1 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope Andreas Premstaller, Ph.D. Biopharmaceutical Operations San Francisco, January 14, 2009 a Novartis company

2 Outline Analytical tools for the characterization of somatropin Comparability studies in the development of Omnitrope : Scope, approaches and execution Case studies of comparability assessments: Biosimilarity Process changes Novel formulations 2 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

3 Analytical tools Comprehensive characterization of somatropin

4 Somatropin, the active ingredient of Omnitrope Recombinant human growth hormone (rhgh) from modified E. coli Chemically identical to pituitary growth hormone, non-glycosylated protein 1 chain of 191 amino acids, 2 disulfide bridges, with 4 antiparallel α-helices C 990 H 1528 N 262 O 300 S 7 Mass Source: PDB ID: 1HGU Chantalat, L. Jones, N.D., Korber, F., Navaza, J., Pavlovsky, A.G. (1995) The crystal-structure of wild-type growth-hormone at 2.5 Angstrom resolution. Protein Pept. Lett. 2: Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

5 Analytical methods to characterize somatropin in comparability studies Molecular parameter Primary structure Method Edman sequencing Peptide map with MS detection Mass Spatial structure (secondary and tertiary) Mass spectrometry: MALDI-TOF, ESI-MS Circular dichroism spectroscopy 1D { 1 H} NMR spectroscopy Polarity Charge Reversed phase chromatography Capillary electrophoresis Isoelectric focusing Ion exchange chromatography Size Size exclusion chromatography Gel electrophoresis Immunological tests Biological activity Immunoblotting In-vivo bioassays Cell proliferation assay 5 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

6 Assessment of identity: Complete sequence coverage by combination of peptide maps AA# T1 T2 T4 T6 FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFLQNPQT G1 G2 G3 G5 G6 AA# T7 T8 T9 T10 SLCFSESIPTPSNREETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANS G7 G T11 T12 T13 T14 T15 LVYGASDSNVYDLLKDLEEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNS G13 AA# T16 T18 T19 T20 T19 HNDDALLKNYGLLYCFRKDMDKVETFLRIVQCRSVEGSCGF G18 G21 G22 Complete sequence coverage is obtained by combination of the detected peptides from digests with trypsin (blue) and Glu-C (orange). 6 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

7 Considerations for analytical methods: Impact of matrix and sample preparation Digestion without buffer exchange Digestion with buffer exchange Somatropin NIBSC 98/574 Somatropin, liquid formulation, ph WHO Omnitrope Product 1 T4 T4+T5 T10 (T6+T7)-S-S-T16 T6-S-S-T16 T1-1 T10-2 T11 Somatropin NIBSC 98/574 Somatropin, liquid formulation, after buffer exchange 04-WHO Product Omnitrope, 1, buffer exchange T4+T5 T10 (T6+T7)-S-S-T16 T6-S-S-T16 T1-1 T10-2 T11 T Sample preparation and analytical methods must be developed to account for differences in sample matrices. 7 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

8 Comparability studies Scope, common elements, differences

9 Comparability studies in the development of Omnitrope Scope Biosimilarity: Comparison to a reference product Process changes: Comparison between a pre- and post-change product Novel formulations: Comparison of different drug product formulations, and assessment of potential differences in the profile of product-related variants 9 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

10 Comparability studies in the development of Omnitrope Common approach Science and knowledge based approach Previous knowledge Risk assessment with respect to potential impact of changes/ differences Identification of Critical Quality Attributes (CQA), Critical Process Parameters (CPP) Focus on analytical methods to monitor CQA s, CPP s Type and extend of studies (physicochemical, biological, preclinical, clinical) is determined in a step-wise approach based on these consideration and the knowledge regarding product and process. 10 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

11 Comparability studies in the development of Omnitrope Execution Setting acceptance criteria: Considering the previous experience with the product at the biosimilar developer and the information in the public domain Biosimilarity: Considering the batch-to-batch variability of the reference product Process changes: Based on the historical performance of the process Differences need to be assessed in appropriate studies and scientifically justified to exclude an impact on efficacy and safety of the product Classification of the product variants into product-related substances or impurities (ICH Q6B) 11 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

12 Comparability studies Biosimilarity

13 Comparability of Omnitrope with the reference product was established at all stages Comparability Clinical efficacy and safety PK/PD Preclinical Biological characterization Physicochemical characterization In a representative indication (growth hormone deficient children) Comparative PK/PD in healthy volunteers 14- day repeat toxicity (rats) local tolerance (rabbits) In-vitro and in-vivo bioassays Comprehensive analysis of protein Complete stand-alone product and target-directed process development Development according to relevant guidelines 13 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

14 Controlling the product profile of somatropin: Detecting potential product-related variants Variant Polarity (RP-HPLC) Charge (CZE, IEF, IEX) Size (SEC) Peptide mapping Deamidated X X - Succinimide X X - - Isoaspartate X - - x Oxidized X - - X Truncated and clipped variants X X - X Trisulfide X - - X Thioether X Aggregates Covalent dimer X - X X X Non-covalent aggregates Dimer, oligomers - - X - X resolved and detected; - not resolvable with this method 14 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

15 A thioether variant of somatropin: Background Disulfide bridges in somatropin: Cys(53)-Cys(165) Cys(182)-Cys(189) Thioether variant: Cys(182)-Cys(189) * IVQCR S S SVEGSCGF IVQCR S SVEGSCGF * Datola, A. et al., ChemMedChem, 2 (2007) Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

16 A thioether variant of somatropin: Detection and identification Detection by peptide mapping Identification by high resolution mass spectrometry Reference product T20-S-S-T21, (M+2H) 2+ Expected mass: Found mass: Omnitrope Somatropin X T20-S-T21 T20-SS-T Time (min) Relative Abundance T20-S-T21, (M+2H) 2+ Expected mass: Found mass: m/z 16 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

17 Phase I PK/PD study program Protocol Design Objective EP2K99PhI S USA EP2K99PhI USA Double-blind, randomised, 2-way cross-over, placebo-controlled study Double-blind, randomised, 2-way cross-over, controlled study Phase I safety study in healthy volunteers to assess the safety, tolerance and pharmacokinetics of Omnitrope lyophilised powder (API Covance). Feasibility study to demonstrate that continuous i.v infusion of octreotide is effective to suppress endogenous GH secretion in healthy subjects Phase I study in healthy volunteers to compare the pharmacokinetics, pharmacodynamics and safety between Omnitrope lyophilised powder (API Covance) and Genotropin No. Subj. Dose 12 5 mg single dose s.c. injection 25 5 mg single dose s.c. injection EP2K00PhI AQ Double-blind, randomised, 2-way cross-over, controlled study Phase I study in healthy volunteers to compare the pharmacokinetics, pharmacodynamics and safety between Omnitrope lyophilised powder (API Covance) and Omnitrope 3.3 mg/ml solution f.i. (API Kundl) 24 5 mg single dose s.c. injection EP Double-blind, randomised, 3-way cross-over, controlled study Phase I study in healthy volunteers to compare the pharmacokinetics, pharmacodynamics and safety between Omnitrope 5mg/ml powder f.s.f.i., Omnitrope 3.3 mg/ml solution f.i. and Genotropin 36 5 mg single dose s.c. injection EP Double-blind, randomised, 3-way cross-over, controlled study Phase I study in healthy volunteers to compare the pharmacokinetics, pharmacodynamics and safety between Omnitrope 5mg/ml powder f.s.f.i., Omnitrope 6.7 mg/ml solution f.i. and Genotropin 36 5 mg single dose s.c. injection 17 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

18 Phase III Clinical Program to confirm comparable safety and efficacy Protocol Design Objective No. Subj. Dose Pivotal efficacy study EP2K99PhIII EP2K00PhIIIFo EP2K00PhIIIAQ part A Randomized, controlled, open, multicenter study in GHD children Controlled, open, multicenter study in GHD children Controlled, open, multicenter study in GHD children Efficacy and safety - Demonstrate therapeutic equivalence between Omnitrope Lyophilized Powder and Genotropin Efficacy and safety - Follow-up study to demonstrate therapeutic equivalence between Omnitrope Lyophilized Powder and Genotropin Efficacy and safety - Demonstrate therapeutic equivalence between Omnitrope Lyophilized Powder and Omnitrope Liquid Formulation 89 Omnitrope Lyophilized Powder vs. Genotropin 0.03 mg/kg daily, sc 86 Omnitrope Lyophilized Powder vs. Genotropin 0.03 mg/kg daily, sc 86 Omnitrope Lyophilized Powder vs. Omnitrope Liquid Formulation 0.03 mg/kg daily, sc EP2K00PhIIIAQ part B Open, multicenter study in GHD children Demonstrate long term safety and efficacy of Omnitrope Liquid Formulation 86 Omnitrope Liquid Formulation 0.03 mg/kg daily, sc Pivotal safety study EP2K02PhIIILyo Open, multicenter study in GHD children Safety and efficacy - Confirmation of low immunogenicity of Omnitrope Lyophilized Powder 51 Omnitrope Lyophilized Powder 0.03 mg/kg daily, sc 18 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

19 Biosimilarity: Comparability with the reference product A biosimilar product is designed to meet the criteria of the reference product with regards to quality, safety and efficacy. Biosimilars development requires complete product and process development plus comparative testing at all relevant levels. The comparability of Omnitrope to the reference product was shown using a comprehensive array of test methods. Pre-clinical and clinical assessment confirmed similarity and clinical equivalence. Reference product Comparative Comparative Targeted development/ Quality by Design Quality Module 3 Physicochemical and biological comparability Comparative preclinical studies Comparative clinical studies Risk Management Plan Biosimilar product 19 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

20 Comparability studies Process changes

21 Comparability assessment for process changes Acceptance criteria for comparability: Based on the historical performance of the process (batch history) and understanding of the product. Data from reference product may be used as supportive data in assessment of the change Approaches: Submission of the entire comparability study Submission of a comparability protocol ahead of implementation of changes Extend of the studies: Varies with extend of change For scale or site changes to a well-controlled and understood process physicochemical and biological data were found appropriate and sufficient 21 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

22 Comparability studies Novel formulations

23 Liquid formulations of somatropin Stable liquid formulation, containing buffer system, tonicity modifying excipients, surface active excipients, antimicrobial preservation agent Lyophilized and liquid formulation after submission to mechanical stress: Particle profile in the subvisible range Particle count 512 channels 0.5 µm 450 µm Intensity (%) Size Distribution by Intensity Dynamic Light Scattering 0.1 nm 1 µm Intensity (%) Size Distribution by Intensity Size (d.nm) Reconstituted powder Size (d.nm) Liquid formulation 23 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

24 Qualification of a liquid formulation of Omnitrope Clinical safety & efficacy PK/PD Preclinical Biological characterization Efficacy and safety of profile of product-related variants confirmed in Phase III studies Equivalence to lyophilized formulation and reference product in Phase I studies Toxicology and local tolerance studies, potency in animal model of fresh and aged product Biological characterization of aged product and product-related variants Physicochemical characterization Stability studies; characterization of aged product and of product-related variants at end of shelf life 24 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

25 Conclusions Assessments of comparability are crucial components in the development of any biopharmaceutical product, and specifically so for biosimilar development. The extend of a comparability study needs to be based on the magnitude of the introduced change and the knowledge of product and process. A set of orthogonal analytical tools is essential to overcome the limitations of single methods and to obtain a complete picture of the quality of the product biosimilar candidate and reference product. The efficacy and safety and the comparability of Omnitrope with the reference product were shown combining data from physicochemical, biological, preclinical and clinical studies. Data from all these levels were used to demonstrate that the liquid formulations of Omnitrope have consistent quality characteristics, and are efficacious and safe formulations of somatropin. 25 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /

26 Acknowledgements Technical team Sabine Fürtinger Bernt Pragl Hansjörg Toll Development concepts and registration Ajaz Hussain Martin Schiestl Ingrid Schwarzenberger Jörg Windisch 26 Comparability assessment in the development of follow-on protein products: Case studies from Omnitrope / AP /