biosimilar development regulatory, analytical, and clinical considerations

BIOSIMILAR DEVELOPMENT: REGULATORY, ANALYTICAL, AND CLINICAL CONSIDERATIONS

SGS LIFE SCIENCE SERVICES WEBINAR

CLINICAL CONSIDERATIONS

April 22, 2015

MEETING REGULATORY ANALYTICAL CHARACTERIZATION EXPECTATIONS


Dr Fiona M GreerGlobal Director, BioPharma Services DevelopmentSGS Life Science Services

AGENDA: MEETING REGULATORY ANALYTICAL 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)

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• Package of analytical tools/ battery of methods

• Strategies for primary and higher order structure

WHY ARE BIOPRODUCTS A CHALLENGE ?

During/ after translation of the oligonucleotidecode 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.

AcetylationAcylationAmidation(deamidation)CarbamylationCarboxylationFormylation

GlycationGlycosylationMethylationMethionine OxidationNorleucinePhosphorylationSulphation

Carbohydrate is a source of product


Carbohydrate is a source of product heterogeneity. Glycoproteins are mixtures of glycoforms i.e. the same polypeptide but different sugars.

EXAMPLE OF COMPLEXITY: ANTIBODY CASE


WHEN IS ANALYTICAL CHARACTERIZATION REQUIRED?


WHAT REGULATIONS COVER PHYSICOCHEMICAL CHARACTERIZATION?

� ICH Topic Q6B “Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products”

� Structural characterization and confirmation1. Amino acid sequence2. Amino acid composition3. Terminal amino acid sequence


3. Terminal amino acid sequence4. Peptide map5. Sulfhydryl group(s) and disulfide bridges6. Carbohydrate structure

� Physicochemical properties1. Molecular weight or size2. Isoform pattern3. Extinction coefficient4. Electrophoretic pattern5. Liquid Chromatographic pattern6. Spectroscopic profiles

POTENTIAL ANALYTICAL TOOLS

� Amino acid 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


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

CASE STUDY: ANTIBODY CHARACTERIZATION

• 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)


and 100% HC)

• Monosaccharide and sialic acid

analysis

• Oligosaccharide population

analysis

• SDS-PAGE analysis

• Circular Dichroism

• Analytical Ultracentrifugation

N-Linked biantennary core fucosylated 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 = +162G2F Mass shift = +324

INTACT MASS MEASUREMENT (MONITORING GLYCOSYLATION)


mAb +2 x G0F

+ 1 x G1FmAb +2 x G1F

mAb +1 x G1F+ 1 x G2F

INTACT MASS COMPARISON OF THREE BIOSIMILAR MABS


PEPTIDE MAPPING WORKFLOW


ANTIBODY ANALYSIS – GENERAL WORKFLOW


SULFHYDRYL GROUP(S) AND DISULFIDE BRIDGES


S S

SH

Disulphide bridgedprotein

E

E

Enzymic/Chemicaldigestion

S S SH

Mixture ofpeptides

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

% I

nte

ns

ity

Voyager Spec #1 MC[BP = 3017.6, 2567]

KTCIVPEVSSVFIFPPKPK

KVTCVVVDISK

252 269

280 289

CHARACTERIZATION OF S-S BRIDGES


SHE

EIdentification by MSFollowed by reductionAnd 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

% In

ten

sit

y

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

% In

ten

sit

y

Voyager Spec #1=>SM5[BP = 1662.4, 7089]1062.6

1988.1VTCVVVDISK280 289

TCIVPEVSSVFIFPPKPK252 269

Reduction

ANALYSIS OF GLYCOSYLATION

COOH2HNS---S

S---S

N-Glycans

O-Glycans

Intact Mass by MALDI or ES MSMonosaccharide Composition Analysis (LC & MS)Reduction Carboxymethylation

COOH2HN

S-CM S-CMS-CMS-CM

Specific Protease Digest


Reductive

elimination

Specific Protease Digest

PNGase F

Sep-pak

0% 20% 40%

Permethylation MALDI,Nanospray-MS/MS & Linkage analysisLC & MS methods

Monosaccharide CompositionGlycan Population ScreeningGlycan Antennary ProfileGlycosylation SiteLinkage Analysis

Native

Glycans

MALDI-MS

Anomeric specific Intermonosaccharide

Linkages Glycan Profile

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


Sample Glycoprotein

ES-MS / MALDI-MS

Quantitative Monosaccharide

Composition

GC-MS

HPAEC-PAD

Quantitative Sialic Acid Content

Derivatised

Glycans

PMAA GC-MS

Inter-monosaccharide

Linkages

Antennary ProfileESI-MS

2AB-LC-MS

Quantitative Glycan profile

Glycopeptides

Qualitative Site-specific Glycosylation

Peptide MappingLC-ES-MS

� MALDI-MS analysis of permethylated N-glycans

MAJOR STRUCTURES OF N-LINKEDOLIGOSACCHARIDES ON IGG1 MABS

Possible structure for the signal at m/z 1836 (G0F)




TIC chromatogramAnnotations based on MS data

OLIGOSACCHARIDE PROFILINGLC- AND MS-BASED METHODS

2-AB labelling and HPLC-FLD for profiling Oligosaccharide population coupled with ESI-MS


Annotations based on MS data

Example of IgG N-glycans

N-acetylglucosamine

Galactose

Mannose

Fucose

N-acetylneuraminic acid

N-glycolylneuraminic acid

BIOPHYSICAL TECHNIQUES FOR HIGHER ORDER STRUCTURE, CONFORMATION AND AGGREGATION

Technique Reports on Advantages Disadvantages

Circular DichroismSecondary/ TertiaryStructure

QuantitativeSensitive to helix content

Formulation buffers can interfere

FTIR Secondary StructureQuantitativeSensitive to sheet contentLess prone to buffer interfence

IntrinsicFluorescence

Local Tertiary StructureSensitivePotential for moderate HTP

Qualitative


Extrinsic Fluorescence Surface hydrophobicitySensitiveEnsemble tertiary structure-no localPotential for moderate HTP

Qualitative

UV-VIS (2ndderivative) Local Tertiary StructureSimultaneous to concentration determinationPotential for moderate HTP

Qualitative

Differential Scanning Calorimetry

Thermal StabilityScreening 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-MALSOligomers/ aggregates

Direct MW determination, rapid analysisMatrix presentHigh shear forces

RE-CAP: ANALYTICAL CHARACTERIZATION DATA FOR 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


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.

Life Science Services Fiona GreerGlobal Director, BioPharma Services Development

SGS M-Scan Ltd Phone: +44 (0) 118 989 6940

THANK YOU FOR YOUR ATTENTION

+ 41 22 739 9548


SGS M-Scan Ltd Phone: +44 (0) 118 989 69402-3 Millars Business Centre, Fax: +44 (0) 118 989 6941Fishponds Close,Wokingham E-mail : [email protected], 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

MEETING THE CHALLENGES OF EARLY PHASE CLINICAL TRIALS WITH BIOSIMILARS


BIOSIMILARS

Annick Van RielDirector of the Clinical Pharmacology Unit SGS Life Science Services

OVERVIEW

� Opportunities

� Regulatory Challenges

� Comparative Clinical Trials

• a Stepwise Approach


• a Stepwise Approach

� Clinical Considerations in phase I

• Operational Challenges

• Operational Solutions

OPPORTUNITIES

� Market for Biosimilars will reach $9,2bn in 2018

� Key drivers• Patent expiry of biologic drugs

– By 2020, some $55bn biologic patents due to expire

• Increasing market demands


• 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

REGULATORY CHALLENGES- REFERENCE PRODUCT

� SELECTION of REFERENCE PRODUCT (RBP)

• Market (Europe / US / Both)

• Indication– Extrapolation across other indications of the RBP will need

scientific justification


� COMPARABILITY in SAFETY AND EFFICACY

• Deviations from RBP (strength, pharmaceutical form, formulation,…) require justification

– Route of administration

– Improved efficacy

– Improved safety

REGULATORY CHALLENGES- DOSSIER

� NON-CLINICAL DEVELOPMENT• ‘Cutting corners’ on EU / FDA guidance on non-clinical

development• Risk, but can be acceptable if product is considered safe

� Chemistry, Manufacturing & Controls (CMC)


� 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/…)

COMPARATIVE CLINICAL TRIALS- A STEPWISE APPROACH

� Clinical evaluation in intended population

� Non-clinical testing

� Analytical comparison to reference product


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.


� CLINICAL DEVELOPMENT

To show comparable safety and efficacy between RBP / SBP

• Detect and explore relevant differences

•


• 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%)



• STEP 1B – Pharmacodynamics– Clear dose response relationship

– Accepted PD markers

– Population in which the possible differences could be best observed


– 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



• STEP 2A – Clinical Efficacy– Designed to prove similar clinical efficacy between RBP/SBP

» Similar treatment effect

» Similar dosage


– 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

CLINICAL TRIAL CONSIDERATIONS IN 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


• 3-arm studies

� Large sample size

� Study design• Cross over or parallel• Half life• Safety• Dose (single / multiple …)• Driven by PD, not PK

CLINICAL TRIAL CONSIDERATIONS IN PHASE I- OPERATIONAL CHALLENGES

� Biosimilar experience

� Bioanalytical knowledge

� Quality – control variability

� Regulatory approval

� Number of cohorts, bed size

� Population -Recruitment


variability

� Pharmacy IMP preparation

� Clinical Conduct

� Sample processing and shipment

Recruitment

� Screening

� Staffing

� Timelines

� Randomized, double-blind, 3-way parallel study to compare PK between X and adalimumab (EU and US sourced).

• PK, safety, tolerability

• Subcutaneous injection

1


• Subcutaneous injection

• Single center, 252 healthy male / female volunteers

� CHALLENGE

• Regulatory– Submission rejected in The Netherlands

• Recruitment

• Screening

• Timelines

� 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


• PK, safety, tolerability, immunogenicity

• IV infusion

• Single center, 147 healthy male volunteers, 120 days

� CHALLENGE • Recruitment

• Screening

• Timelines

CLINICAL TRIAL CONSIDERATIONS IN 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


– CA comments

» Clarification on manufacturing sites

» Recommendation on in vitro assay in non-clinical program

– IRB/EC comments

» None received


� 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


• Continuous building of CPU healthy volunteer database to expand biologic-naïve population, due to sponsor preferences.


� RECRUITMENT and SCREENING SOLUTIONS• Contracted Call Centre

• Advertisement

• Sources

• Sufficient volunteer fee, linked to duration

• Start immediately after EC approval


• 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


� 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


• Line-up for screening

• Continuous inclusion

• Fast data processing– Continuous data processing (on-site and data management)

– Fast database lock due to flexible teams

CONCLUSION

� Biosimilar Clinical Development in phase I has challenges

• High Clinical cost, resource and time– Plan complete programme, not 1 study

• No “one-size fits all” approach


• 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

QUESTIONS

Speakers:

Nadine M. Ritter, Ph.D.Global Biotech [email protected]

Dr. Fiona M. GreerSGS Life Science Services [email protected]


[email protected]

Annick Van RielSGS Life Science [email protected]

For additional information visit:

WWW.SGS.COM/BIOSIMILARS

Thank you for attending this event.

biosimilar development regulatory, analytical, and clinical considerations

Healthcare