Download as PDF, PPTX
![S S
SH
Disulphide bridged
protein
E
E
Enzymic/Chemical
digestion
S S SH
Mixture of
peptides
2971.0 2989.6 3008.2 3026.8 3045.4 3064.0
Mass (m/z)
0
2567.0
0
10
20
30
40
50
60
70
80
90
100
%Intensity
Voyager Spec #1 MC[BP = 3017.6, 2567]
KTCIVPEVSSVFIFPPKPK
KVTCVVVDISK
252 269
280 289
CHARACTERIZATION OF S-S BRIDGES
15© SGS SA 2015 ALL RIGHTS RESERVED
SH
E
E
Identification by MS
Followed by reduction
And further MS
1154.0 1169.4 1184.8 1200.2 1215.6 1231.0
Mass (m/z)
1122.8
20
30
40
50
60
70
80
90
100
%Intensity
Voyager Spec #1=>SM5[BP = 1662.4, 7089]
1955 1970 1985 2000 2015 2030
Mass (m/z)
754.3
10
20
30
40
50
60
70
80
90
100
%Intensity
Voyager Spec #1=>SM5[BP = 1662.4, 7089]
1062.6
1988.1
VTCVVVDISK
280 289
TCIVPEVSSVFIFPPKPK
252 269
Reduction](https://image.slidesharecdn.com/sgs-lss-biosimilarswebinaranalyticalandclinicalconsiderations-en-15-150512142519-lva1-app6891/85/Biosimilar-Development-Regulatory-Analytical-and-Clinical-Considerations-15-320.jpg)
Uploaded bySGS
Biosimilar Development Regulatory, Analytical, and Clinical Considerations
The document discusses the complex landscape of biosimilar development, highlighting regulatory, analytical, and clinical considerations essential for successful product characterization and comparability. Key challenges include the need for thorough analytical techniques to characterize biologics and establish a quality target product profile, alongside stringent regulatory requirements for demonstrating biosimilarity. The document outlines a stepwise approach for clinical trials, emphasizing the importance of comparative studies in assessing safety and efficacy between reference and biosimilar products.
More Related Content
Similar to Biosimilar Development Regulatory, Analytical, and Clinical Considerations
Biosimilar Development Regulatory, Analytical, and Clinical Considerations
- 1. BIOSIMILAR DEVELOPMENT: REGULATORY, ANALYTICAL,AND CLINICAL CONSIDERATIONS SGS LIFE SCIENCE SERVICES WEBINAR CLINICAL CONSIDERATIONS April 22, 2015
- 2. MEETING REGULATORY ANALYTICAL CHARACTERIZATIONEXPECTATIONS BIOSIMILAR DEVELOPMENT: REGULATORY, ANALYTICAL, AND CLINICAL CONSIDERATIONS Dr Fiona M Greer Global Director, BioPharma Services Development SGS Life Science Services
- 3. AGENDA: MEETING REGULATORYANALYTICAL CHARACTERIZATION EXPECTATIONS What are the challenges in characterizing complex protein/glycoprotein products? When is analytical characterization required? Which techniques, old & new, are suitable • Establishing the Quality Target Product Profile (QTPP) 3© SGS SA 2015 ALL RIGHTS RESERVED • Package of analytical tools/ battery of methods • Strategies for primary and higher order structure
- 4. WHY ARE BIOPRODUCTSA CHALLENGE ? During/ after translation of the oligonucleotide code into an AA sequence, processing events occur to confer biological activity. These Co- and Post-Translational events change the primary structure but are NOT predictable from the gene sequence. So it is essential to study the expressed protein products not the genes. Acetylation Acylation Amidation (deamidation) Carbamylation Carboxylation Formylation Glycation Glycosylation Methylation Methionine Oxidation Norleucine Phosphorylation Sulphation Carbohydrate is a source of product 4© SGS SA 2015 ALL RIGHTS RESERVED Carbohydrate is a source of product heterogeneity. Glycoproteins are mixtures of glycoforms i.e. the same polypeptide but different sugars.
- 5. EXAMPLE OF COMPLEXITY:ANTIBODY CASE 5© SGS SA 2015 ALL RIGHTS RESERVED
- 6. WHEN IS ANALYTICALCHARACTERIZATION REQUIRED? 6© SGS SA 2015 ALL RIGHTS RESERVED
- 7. WHAT REGULATIONS COVER PHYSICOCHEMICALCHARACTERIZATION? ICH Topic Q6B “Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products” Structural characterization and confirmation 1. Amino acid sequence 2. Amino acid composition 3. Terminal amino acid sequence 7© SGS SA 2015 ALL RIGHTS RESERVED 3. Terminal amino acid sequence 4. Peptide map 5. Sulfhydryl group(s) and disulfide bridges 6. Carbohydrate structure Physicochemical properties 1. Molecular weight or size 2. Isoform pattern 3. Extinction coefficient 4. Electrophoretic pattern 5. Liquid Chromatographic pattern 6. Spectroscopic profiles
- 8. POTENTIAL ANALYTICAL TOOLS Aminoacid sequence and modifications: MS, peptide mapping, chromatography Glycosylation: Anion exchange, enzymatic digestion, peptide mapping, CE, MS Folding: MS S-S bridge determination, calorimetry, HDX and ion mobility MS, NMR, circular dichroism, Fourier transform spectroscopy, fluorescence 8© SGS SA 2015 ALL RIGHTS RESERVED spectroscopy, fluorescence PEGylation & isomers: chromatography, peptide mapping Aggregation: Analytical ultracentrifugation, size-exclusion chromatography SEC-MALS, field flow fractionation A4F, light scattering DLS, microscopy, TEM Proteolysis: electrophoresis, chromatography, MS Impurities: proteomics, immunoassays, metal & solvents analysis Subunit interactions: chromatography, ion mobility MS Heterogeneity of size, charge, hydrophobicity: Chromatography; gel & capillary electrophoresis, light scattering, IM-MS, CESI-MS
- 9. CASE STUDY: ANTIBODYCHARACTERIZATION • Mass spectrometry of intact protein and released L &H chains • Amino Acid Composition Analysis • N- and C-terminal sequencing • Peptide “MAPPING” Analysis (Sequence coverage: 100% LC and 100% HC) 9© SGS SA 2015 ALL RIGHTS RESERVED and 100% HC) • Monosaccharide and sialic acid analysis • Oligosaccharide population analysis • SDS-PAGE analysis • Circular Dichroism • Analytical Ultracentrifugation
- 10. N-Linked biantennary corefucosylated with varying number of galactose residues Fuc Man – GlcNAc Asn - GlcNAc-GlcNAc- Man Man - GlcNAc - Gal - Gal mAb +1 x G0F + 1 x G1F mAb +2 x G1F G0F Mass shift = +1444 G1F Mass shift = +162 G2F Mass shift = +324 INTACT MASS MEASUREMENT (MONITORING GLYCOSYLATION) 10© SGS SA 2015 ALL RIGHTS RESERVED mAb +2 x G0F + 1 x G1F mAb +2 x G1F mAb +1 x G1F + 1 x G2F
- 11. INTACT MASS COMPARISONOF THREE BIOSIMILAR MABS 11© SGS SA 2015 ALL RIGHTS RESERVED
- 12. PEPTIDE MAPPING WORKFLOW 12©SGS SA 2015 ALL RIGHTS RESERVED
- 13. ANTIBODY ANALYSIS –GENERAL WORKFLOW 13© SGS SA 2015 ALL RIGHTS RESERVED
- 14. SULFHYDRYL GROUP(S) ANDDISULFIDE BRIDGES 14© SGS SA 2015 ALL RIGHTS RESERVED
- 15. S S SH Disulphide bridged protein E E Enzymic/Chemical digestion SS SH Mixture of peptides 2971.0 2989.6 3008.2 3026.8 3045.4 3064.0 Mass (m/z) 0 2567.0 0 10 20 30 40 50 60 70 80 90 100 %Intensity Voyager Spec #1 MC[BP = 3017.6, 2567] KTCIVPEVSSVFIFPPKPK KVTCVVVDISK 252 269 280 289 CHARACTERIZATION OF S-S BRIDGES 15© SGS SA 2015 ALL RIGHTS RESERVED SH E E Identification by MS Followed by reduction And further MS 1154.0 1169.4 1184.8 1200.2 1215.6 1231.0 Mass (m/z) 1122.8 20 30 40 50 60 70 80 90 100 %Intensity Voyager Spec #1=>SM5[BP = 1662.4, 7089] 1955 1970 1985 2000 2015 2030 Mass (m/z) 754.3 10 20 30 40 50 60 70 80 90 100 %Intensity Voyager Spec #1=>SM5[BP = 1662.4, 7089] 1062.6 1988.1 VTCVVVDISK 280 289 TCIVPEVSSVFIFPPKPK 252 269 Reduction
- 16. ANALYSIS OF GLYCOSYLATION COOH2HN S---S S---S N-Glycans O-Glycans IntactMass by MALDI or ES MS Monosaccharide Composition Analysis (LC & MS)Reduction Carboxymethylation COOH2HN S-CM S-CMS-CMS-CM Specific Protease Digest 16© SGS SA 2015 ALL RIGHTS RESERVED Reductive elimination Specific Protease Digest PNGase F Sep-pak 0% 20% 40% Permethylation MALDI, Nanospray-MS/MS & Linkage analysis LC & MS methods Monosaccharide Composition Glycan Population Screening Glycan Antennary Profile Glycosylation Site Linkage Analysis
- 17. Native Glycans MALDI-MS Anomeric specific Intermonosaccharide Linkages GlycanProfile HPAEC-PAD Intact mass vs. Deglycosylated ES-MS / MALDI-MS Heterogeneity & Extent of Glycosylation Derivatised Glycan Composition Glycan Sequence MALDI-MS MALDI-MS/MS Antennary GLYCOMIC / GLYCOPROTEOMIC WORKFLOW 17© SGS SA 2015 ALL RIGHTS RESERVED SampleGlycoprotein ES-MS / MALDI-MS Quantitative Monosaccharide Composition GC-MS HPAEC-PAD Quantitative Sialic Acid Content Derivatised Glycans PMAA GC-MS Inter- monosaccharide Linkages Antennary Profile ESI-MS 2AB-LC-MS Quantitative Glycan profile Glycopeptides Qualitative Site-specific Glycosylation Peptide Mapping LC-ES-MS
- 18. MALDI-MS analysis ofpermethylated N-glycans MAJOR STRUCTURES OF N-LINKED OLIGOSACCHARIDES ON IGG1 MABS Possible structure for the signal at m/z 1836 (G0F) 18© SGS SA 2015 ALL RIGHTS RESERVED Possible structure for the signal at m/z 2040 (G1F) Possible structure for the signal at m/z 2244 (G2F)
- 19. TIC chromatogram Annotations basedon MS data OLIGOSACCHARIDE PROFILING LC- AND MS-BASED METHODS 2-AB labelling and HPLC-FLD for profiling Oligosaccharide population coupled with ESI-MS 19© SGS SA 2015 ALL RIGHTS RESERVED Annotations based on MS data Example of IgG N-glycans N-acetylglucosamine Galactose Mannose Fucose N-acetylneuraminic acid N-glycolylneuraminic acid
- 20. BIOPHYSICAL TECHNIQUES FORHIGHER ORDER STRUCTURE, CONFORMATION AND AGGREGATION Technique Reports on Advantages Disadvantages Circular Dichroism Secondary/ Tertiary Structure Quantitative Sensitive to helix content Formulation buffers can interfere FTIR Secondary Structure Quantitative Sensitive to sheet content Less prone to buffer interfence Intrinsic Fluorescence Local Tertiary Structure Sensitive Potential for moderate HTP Qualitative 20© SGS SA 2015 ALL RIGHTS RESERVED Extrinsic Fluorescence Surface hydrophobicity Sensitive Ensemble tertiary structure-no local Potential for moderate HTP Qualitative UV-VIS (2ndderivative) Local Tertiary Structure Simultaneous to concentration determination Potential for moderate HTP Qualitative Differential Scanning Calorimetry Thermal Stability Screening method for formulation (HTP) Qualitative SV-AUC Oligomers/ aggregates Matrix free, quantitative, resolution Slow, DLS HMW aggregates Sensitivity, moderate for HTP Poor resolution, qualitative SEC-MALS Oligomers/ aggregates Direct MW determination, rapid analysis Matrix present High shear forces
- 21. RE-CAP: ANALYTICAL CHARACTERIZATION DATAFOR BIOSIMILARS Development of a Biosimilar requires comprehensive physicochemical structural characterization at MANY stages. Initially, batches of originator are studied to determine the exact protein sequence, PTMs and variability of quality attributes. These data form the Quality Target Product Profile (QTPP). MS techniques are applicable at all stages of development, but essential for determination of originator sequence. Advances in 21© SGS SA 2015 ALL RIGHTS RESERVED essential for determination of originator sequence. Advances in instrumentation and Proteomic/Glycomic strategies enable rapid identification of QTPP including PTMs. At early stage, characterization surveys may help to guide choice of an appropriate cell line. Build similarity concept from start. Various regulatory guidelines then require side-by-side comparative data to demonstrate “Biosimilarity”. MS alone is not enough. Multiple orthogonal analytical methods are used to define “fingerprint” comparison. Increasing importance on HOS techniques to link with biological activity.
- 22. Life Science ServicesFiona Greer Global Director, BioPharma Services Development SGS M-Scan Ltd Phone: +44 (0) 118 989 6940 THANK YOU FOR YOUR ATTENTION + 41 22 739 9548 22© SGS SA 2015 ALL RIGHTS RESERVED SGS M-Scan Ltd Phone: +44 (0) 118 989 6940 2-3 Millars Business Centre, Fax: +44 (0) 118 989 6941 Fishponds Close, Wokingham E-mail : fiona.greer@sgs.com Berkshire, RG41 2TZ, UK Web : www.sgs.com/biosimilars + 41 22 739 9548 + 1 866 SGS 5003 + 65 637 90 111 + 33 1 53 78 18 79 + 1 877 677 2667 + 33 1 41 24 87 87
- 23. MEETING THE CHALLENGESOF EARLY PHASE CLINICAL TRIALS WITH BIOSIMILARS BIOSIMILAR DEVELOPMENT: REGULATORY, ANALYTICAL, AND CLINICAL CONSIDERATIONS BIOSIMILARS Annick Van Riel Director of the Clinical Pharmacology Unit SGS Life Science Services
- 24. OVERVIEW Opportunities Regulatory Challenges Comparative ClinicalTrials • a Stepwise Approach 24© SGS SA 2015 ALL RIGHTS RESERVED • a Stepwise Approach Clinical Considerations in phase I • Operational Challenges • Operational Solutions
- 25. OPPORTUNITIES Market for Biosimilarswill reach $9,2bn in 2018 Key drivers • Patent expiry of biologic drugs – By 2020, some $55bn biologic patents due to expire • Increasing market demands 25© SGS SA 2015 ALL RIGHTS RESERVED • Increasing market demands – Aging population – Health awareness – Affordability and insurance coverage • Increasing healthcare cost • Legislations (US, EU, Asia) • Competitive landscape Affordable, safe, efficacious biological drugs
- 26. REGULATORY CHALLENGES - REFERENCEPRODUCT SELECTION of REFERENCE PRODUCT (RBP) • Market (Europe / US / Both) • Indication – Extrapolation across other indications of the RBP will need scientific justification 26© SGS SA 2015 ALL RIGHTS RESERVED COMPARABILITY in SAFETY AND EFFICACY • Deviations from RBP (strength, pharmaceutical form, formulation,…) require justification – Route of administration – Improved efficacy – Improved safety
- 27. REGULATORY CHALLENGES - DOSSIER NON-CLINICALDEVELOPMENT • ‘Cutting corners’ on EU / FDA guidance on non-clinical development • Risk, but can be acceptable if product is considered safe Chemistry, Manufacturing & Controls (CMC) 27© SGS SA 2015 ALL RIGHTS RESERVED Chemistry, Manufacturing & Controls (CMC) • CMC document should clearly demonstrate similarity • Comparability exercise well presented PERCEIVED RISKS of biosimilars • Supportive EC that understands what a biosimilar is • Scientific Advice meeting with Regulatory Authority (FDA/EMA/…)
- 28. COMPARATIVE CLINICAL TRIALS -A STEPWISE APPROACH Clinical evaluation in intended population Non-clinical testing Analytical comparison to reference product 28© SGS SA 2015 ALL RIGHTS RESERVED to reference product • Structure • Function Each step supported by preceding one. Published in: Mark A Socinski; Giuseppe Curigliano; Ira Jacobs; Barry Gumbiner; Judith MacDonald; Dolca Thomas; mAbs 2015, 7, 286-293.
- 29. COMPARATIVE CLINICAL TRIALS -A STEPWISE APPROACH CLINICAL DEVELOPMENT To show comparable safety and efficacy between RBP / SBP • Detect and explore relevant differences • 29© SGS SA 2015 ALL RIGHTS RESERVED • Specific indication • 3-arm trial design • STEP 1A – Pharmacokinetics – Homogenous population, eg healthy volunteers – Supportive PK data from patients – Selected, single dose » Except where dose / time dependent PK – Similarity acceptance range should be pre-defined (80-125%)
- 30. COMPARATIVE CLINICAL TRIALS -A STEPWISE APPROACH CLINICAL DEVELOPMENT • STEP 1B – Pharmacodynamics – Clear dose response relationship – Accepted PD markers – Population in which the possible differences could be best observed 30© SGS SA 2015 ALL RIGHTS RESERVED – Population in which the possible differences could be best observed • Comparative PK / PD trial may be sufficient – PK of RBP well characterized – Sufficient knowledge on PD properties / action mechanism – One PD marker is linked to efficacy – Dose/exposure and response/efficacy of RBP is well established To be discussed with EMA / FDA during Scientific Advise
- 31. COMPARATIVE CLINICAL TRIALS -A STEPWISE APPROACH CLINICAL DEVELOPMENT • STEP 2A – Clinical Efficacy – Designed to prove similar clinical efficacy between RBP/SBP » Similar treatment effect » Similar dosage 31© SGS SA 2015 ALL RIGHTS RESERVED – Similarity acceptance range should be pre-defined – Choose most sensitive population – Extrapolation of indication possible based on overall evidence of biosimilarity • STEP 2B – Clinical Safety – Type, severity, frequency of ADRs between RBP/SBP – Assessment of immunogenicity – Pharmacovigilance and risk management plan
- 32. CLINICAL TRIAL CONSIDERATIONSIN PHASE I - OPERATIONAL CHALLENGES Comparability – NOT Characterization PK and PD endpoints (primary / secondary) Large inter and intra subject variability Bridging with US and EU reference • 3-arm studies 32© SGS SA 2015 ALL RIGHTS RESERVED • 3-arm studies Large sample size Study design • Cross over or parallel • Half life • Safety • Dose (single / multiple …) • Driven by PD, not PK
- 33. CLINICAL TRIAL CONSIDERATIONSIN PHASE I - OPERATIONAL CHALLENGES Biosimilar experience Bioanalytical knowledge Quality – control variability Regulatory approval Number of cohorts, bed size Population - Recruitment 33© SGS SA 2015 ALL RIGHTS RESERVED variability Pharmacy IMP preparation Clinical Conduct Sample processing and shipment Recruitment Screening Staffing Timelines
- 34. Randomized, double-blind, 3-wayparallel study to compare PK between X and adalimumab (EU and US sourced). • PK, safety, tolerability • Subcutaneous injection 1 34© SGS SA 2015 ALL RIGHTS RESERVED • Subcutaneous injection • Single center, 252 healthy male / female volunteers CHALLENGE • Regulatory – Submission rejected in The Netherlands • Recruitment • Screening • Timelines
- 35. Randomised, double-blind, three-arm,parallel group, single dose study to compare the PK, safety, tolerability and immunogenicity of X and reference product (EU and US sourced) in healthy male subjects. • PK, safety, tolerability, immunogenicity 2 35© SGS SA 2015 ALL RIGHTS RESERVED • PK, safety, tolerability, immunogenicity • IV infusion • Single center, 147 healthy male volunteers, 120 days CHALLENGE • Recruitment • Screening • Timelines
- 36. CLINICAL TRIAL CONSIDERATIONSIN PHASE I - OPERATIONAL SOLUTIONS REGULATORY SOLUTIONS • CMC file extensively reworked, including head to head comparison of analytical data • Recent experience in Belgium (15 biosimilar trials) – 100% approval rate for biosimilar studies 36© SGS SA 2015 ALL RIGHTS RESERVED – CA comments » Clarification on manufacturing sites » Recommendation on in vitro assay in non-clinical program – IRB/EC comments » None received
- 37. CLINICAL TRIAL CONSIDERATIONSIN PHASE I - OPERATIONAL SOLUTIONS POPULATION SOLUTIONS • Sponsor preference = biosimilar naïve subjects • Belgium CA allow phase I biosimilar studies to be performed in non-biologic naïve subjects. – Appropriate wash-out times to be observed 37© SGS SA 2015 ALL RIGHTS RESERVED • Continuous building of CPU healthy volunteer database to expand biologic-naïve population, due to sponsor preferences.
- 38. CLINICAL TRIAL CONSIDERATIONSIN PHASE I - OPERATIONAL SOLUTIONS RECRUITMENT and SCREENING SOLUTIONS • Contracted Call Centre • Advertisement • Sources • Sufficient volunteer fee, linked to duration • Start immediately after EC approval 38© SGS SA 2015 ALL RIGHTS RESERVED • Start immediately after EC approval • Line up – Subject availability for multiple groups – Reserve subjects (4 per group) • Plan – Optional screening dates scheduled up front – Optional groups – Protocol to allow for re-screening of subjects • Communicate – Close liaison with investigator and sponsor
- 39. CLINICAL TRIAL CONSIDERATIONSIN PHASE I - OPERATIONAL SOLUTIONS TIMELINE SOLUTIONS • Short approval timelines in Belgium – 15 day HA/IRB(EC) review timelines – Generic screening before CA approval High number of subjects on short # weeks • Line-up for screening 39© SGS SA 2015 ALL RIGHTS RESERVED • Line-up for screening • Continuous inclusion • Fast data processing – Continuous data processing (on-site and data management) – Fast database lock due to flexible teams
- 40. CONCLUSION Biosimilar Clinical Developmentin phase I has challenges • High Clinical cost, resource and time – Plan complete programme, not 1 study • No “one-size fits all” approach 40© SGS SA 2015 ALL RIGHTS RESERVED • No “one-size fits all” approach • Manufacturing complex and challenging • Regulatory requirements – Consult regulatory bodies at planning stage • Phase I site experience – Subject recruitment – timelines
- 41. QUESTIONS Speakers: Nadine M. Ritter,Ph.D. Global Biotech Experts Nadine.Ritter@GlobalBiotechExperts.com Dr. Fiona M. Greer SGS Life Science Services fiona.greer@sgs.com 41© SGS SA 2015 ALL RIGHTS RESERVED fiona.greer@sgs.com Annick Van Riel SGS Life Science Services annick.vanriel@sgs.com For additional information visit: WWW.SGS.COM/BIOSIMILARS Thank you for attending this event.