Regulatory considerations on higher order structure ... · An agency of the European Union Regulatory considerations on higher order structure determination and evaluation –an EU

An agency of the European Union

Regulatory considerations on higher order structure determination and evaluation – an EU perspective

Dr Veronika JekerleQuality OfficeSpecialised Scientific DisciplinesEuropean Medicines Agency

Presented at CASSS-HOS 11-14 April 2016, Long Beach, USA

Contents

Higher order characterisation

• Regulatory expectations

• Observations (case studies)

Considerations on

• Comparability

• Biosimilarity

• Immunogenicity

Update from the EMA (News/Events/Guidance)

Regulatory considerations on HOS – EU perspective © V Jekerle 1

Regulatory expectations

HOS characterisation

Characterisation, identity and consistency

Structure-function relationships (HOS/Potency)

• Functional domains: specificity for binding & binding epitopes

Formulation / PK / cellular update

Stability

Safety features

• Toxicity

• Immunogenicity

• Loss of efficacy

2

Biosimilar development

Comparability assessment

New molecule characterisation

and control

Regulatory considerations on HOS – EU perspective © V Jekerle


Guidelines

3

Note for Guidance on Specifications: Test Procedures and

Acceptance Criteria for

Biotechnological/Biological Products (ICH Q 6 B)

• Composition, physical properties, and primary structure

• higher-order structure

• structural heterogeneity due to biosynthetic processes mixture of post-translationally

modified forms (e.g. glycoforms)

• Biological activity (animal-based, cell culture, biochemical assays, ligand/receptor binding)

• Immunological properties (binding assays)



Guidelines

4

Guideline on development, production, characterisation and

specifications for monoclonal antibodies and related products

(EMEA/CHMP/BWP/157653/2007)

• Amino acid sequence

• Variability of N- and C- terminal amino-acid sequences

• Free sulphydryl groups and disulfide bridges; disulfide bridge integrity mismatch

• Carbohydrate content (neutral sugars, amino sugars and sialic acids)

• Structure of carbohydrate chains, oligosaccharide pattern (antennary profile)

• Glycosylation site(s) and occupancy

• Additional glycosylation site(s) in heavy chains, their presence / absence

• Glycan structures (i.e. degree of mannosylation, galactosylation, fucosylation and sialylation)

• Distribution main glycan structures (often G0, G1 and G2)

• Immunological properties (binding assays)

• Biological activity (defined biological effect)


also relevant

for AB-Drug

conjugates


Guidelines

5

Guideline on similar biological medicinal products containing

biotechnology-derived proteins as active substance: quality issues

Rev. 2014 (EMA/CHMP/BWP/247713/2012)

• Amino acid sequence (same RMP)

• N- and C-terminal amino acid sequences, free SH groups and disulfide bridges

• Modifications/truncations quantified

• Micro-heterogeneous pattern (e.g. C-terminal lysine variability).

• Post-translational modifications (e.g. glycosylation, oxidation, deamidation, truncation)

• Carbohydrate structures (i.e. overall glycan profile, site-specific glycosylation patterns, site occupancy)

• Presence of glycosylation structures / variants not observed in RMP

(� non-human structures (linkages, sequences or sugars))

• Biological activity

• Immunochemical properties


HOS methods – what do Regulators expect?

key points

• Guidance does not prescribe a specific technique

Feasibility /time lag between technique development and implementation as QC

method

Scientific advice on appropriateness of test method(s)

(http://www.ema.europa.eu/ema/index.jsp?curl=pages/regulation/general/general_content_000049.jsp&mid=WC0b01ac05800229b9)

• Combination of physicochemical and biological methods

• Prior knowledge/literature on product class may

reduce burden on testing panel (focus on essential elements)

Benchmark of methods?

Circular dichroism/intrinsic fluorescence/disulfide mapping/ELISA/Functional

assay (?)6 Regulatory considerations on HOS – EU perspective © V Jekerle

What do we see in our dossiers?

Monoclonal antibodies (1997 – 2016)

4 Innovators (cancer & autoimmune indication)

2 Biosimilars

1 AB-drug conjugate

Physico-chemical methods Biological methods

Characterisation Characterisation

Active substance specifications Active substance specifications

Finished product specifications Finished product specifications

Note: methods not relevant to structure (e.g. Bioburden, sterility etc. not included)7


Innovators - physical-chemical methods

Presentation title (to edit, click Insert > Header & Footer)8

Innovator age (4 oldest - 1 youngest)

4 3 2 1

Characterisation

AS specifications

FP specifications


9

Innovator age (4 oldest - 1 youngest)

4 3 2 1

Characterisation

AS specifications

FP specifications

Innovators - biological methods


10

Biosimilar example / function

Example 1 Example 2

Characterisation

AS specifications

FP specifications

Biosimilar - physical-chemical methods


11

Biosimilar example / function

Example 1 Example 2

Characterisation

AS specifications

FP specifications

Biosimilar - biological methods


Observations

Degree of characterisation does not always increase in newer applications

Degree of characterisation is dependent on indication / mechanism of action

Degree of characterisation is dependent on applicant

Applications using QbD principles employ a significantly larger test panel than

comparable indications ( can increased product knowledge be leveraged post-

marketing?)

Biosimilar applications employ a very extensive characterisation panel (Biosimilarity

exercise), which is not translated into more extensive AS/FP specifications

AS and FP specifications rely on 1 or 2 representative bioassays across the board

(even though a much larger number of assays have been established for

characterisation)

12 Regulatory considerations on HOS – EU perspective © V Jekerle

What issues did the CHMP raise?

Innovators (some examples)

• use of FTIR spectroscopy alone for HOS, request to consider also CD

• not clear whether all relevant receptor bindings have been studied (effects on

S/E sufficiently characterised)?

• Acceptance limits of acidic variants (IE-HPLC) are only based on 1 clinical

batch, which is not accepted. Limits should be based on all batch results.

• Method validation (CE-SDS method) with respect to linearity and accuracy

• Cell-based potency assay lacks precision, ELISA assay should be added to

AS/FP release specifications

• Comparison between two methods has not been shown (ADCC cell-based

bioassay and ELISA), both to be used as specifications


What issues did the CHMP raise?

Biosimilars (some examples)

• ‘major’: - demonstration of biosimilarity not shown on the basis of observed

different receptor binding, ADCC activity and small differences in S/E observed

• ‘major’: - dataset not sufficient to conclude on biosimilarity. Additional

justification /data needed to justify that observed differences (in gycosylation)

do not impact S/E

• no specification have been set for charge variants and size variants (other than

HMW)

• no specifications set for glycosylation. Means to monitor glycosylation and

glycan structures at AS release are requested


Antibody-drug conjugate

Focus on size, conjugation and levels of unconjugated mAB (impact of conjugation)

Biological methods!


AB-drug conjugate Physical-chemical methods Biological methods

Characterisation

AS specifications

FP specifications

Considerations on

Comparability, Biosimilarity &

Immunogenicity


molecular heterogeneity to determine that HOS (secondary,

tertiary, and quaternary structure) is maintained

relevant biological activity assay

quality attributes of pre‐change and post‐change products are not

necessarily identical but highly similar

differences in quality attributes have no adverse impact on safety or

efficacy

combination of physico-chemical testing, biological assays ±

nonclinical and clinical data

Comparability

General principles (ICH Q5E)


Evaluation of

quality attributes

no differences differences

“highly similar”

No impact on

S/E


but doubt on

analytical

procedure


no impact on

S/E expected

based on

experience &

relevant data


impact on

S/E cannot be

excluded

significant

differences

comparable

non-clinical/

clinical studiesnon-clinical /

clinical studies

not

comparable

(outside

of scope)

comparable?

ICH Q5E Comparability of Biotechnological/Biological Products

Outcomes HOS


nonclinical

development

clinical development

phase I II IIIpostmarketing

Value of preclinical

data ‘lower’

Focus on product

characterisation &

product definition

Comparability exercise

Clinical bridging study

may be needed

Post-marketing follow-up

necessary?

Comparability exercise

to link earlier clinical data

to product

More substantiated if

during pivotal trial

comparability data

Comparability

Importance of comparability data (ICH Q5E)

ideally no changes


Biosimilarity


21

Dossier requirements for Biosimilars

CTD Module Originator Biosimilar

3

Cross reference

4

5

Cross reference

Cross reference –

class specific

Safety and Efficacy

Integrated Comparability Exercise –

product specific

Quality, Safety and Efficacy

Quality

Non-Clinical

Clinical


22

Biosimilar comparability vs. Comparability

Comparability Biosimilar comparability

Material used Active substance (+ finished product)

Finished product (representative of active substance)

Comparator Earlier stages of same product (clinical development / authorised)

EU-authorised reference product (non-EU if NC/C data from non-EU RP)

Reference In-House reference standard (or Int. Std. if available)

Reference product (in particular the Quality target product profile (QTPP))

Approach Batch data and historic data, specifications

Side-by-side comparison, target product profile

Differences May be expected (depending on complexity of proposed change)

Differences expected (i.e. new process new MCB)

Strategy Analytical comparability (QC and characterisation data)

Stepwise approach – foundation on Quality data. Focus on biological activity

Clinical data Rarely needed Always requested (always needed ?)


23

Immunogenicity

Multidisciplinary approach

Protein

sequence

HOS

Formulation

Packaging

Impurities

Administration

Dose

Frequency

Co-Medication

Co-morbidity

Age, Genotype

NutritionManufacturing

processRegulatory considerations on HOS – EU perspective © V Jekerle

24

‘Biotechnology-derived analogues to human endogenous proteins may

trigger an immune response due to variations in the amino acid

sequence or changes to the protein structure as a result of post-

translational modifications or other changes during all steps of the

manufacturing process, storage and administration.’

‘Glycosylation can influence both the physico-chemical and biological properties of a protein. The

presence and structure of carbohydrate moieties may have both a direct or indirect impact on the

immunogenicity of therapeutic proteins; the glycan can induce an immune response itself (e.g.

glycans of non-human origin), or its presence may affect the conformation of the protein in such

a way that the protein becomes immunogenic.

However, pegylation and glycosylation may also decrease immunogenicity by shielding the

immunogenic epitopes, while maintaining the native conformation of the protein.’


• 31 Jan 2016 (end of public consultation)

• Finalisation stage

Immunogenicity WS, EMA, 9 March 2016, S. Gross25

Without Glycan

Deglycosylation

Without core Fucose

GlcNAc/Mannose (G0)

α1,3Galactose

Galactose

N-glycoylneuraminic acid

N-acetylneuraminic acid

(Sialic acid)

No ADCC activity

Increased immunogenicity

increases ADCC activity

(Mannose binding Lectin)

and Complement activation

higher antigenicity

FcRn binding, CDC activity

higher antigenicity

decreases ADCC activity,

anti-inflammatory

Effects induced by Carbohydrate to IgG Function

from Chung et al., N Engl J Med 2008;358:1109-17 / S. Gross Immunogenicity WS March 2016 26

Cetuximab-Induced Anaphylaxis and IgE Specific for Galactose-α-

1,3-Galactose

25/76 cetuximab-treated subjects had hypersensitivity reaction to the drug

antibodies specific to galactose-α-1,3-galactose.

Product-related

factors

MoA

Structure

Aggregates

Formulation

Purity

Packaging

Storage/Handling…

Risk-based thinking

27

Patient-related factors

Disease

Genetic

Medication

Route of administration

Dose/Dosing regime

…

Risk

Probability

=

x

Consequences

Adapted from C. Schneider (Immunogenicity WS March 2016)

Data/knowledge driven

Impact on Benefit-Risk

Therapeutic benefit

Alternative therapies

Ability to manage risk

Risk mitigation

Follow-up/monitoring

Regular review

during lifecycle

Presentation title (to edit, click Insert> Header and Footer)28

Update from the EMA

News/Events/Guidance

Immunogenicity Workshop 9 March 2016


http://www.ema.europa.eu/ema/index.jsp?curl=pages/news_and_events/events/2016/01/event_detail_001257.jsp&mid=WC0b01ac058004d5c3

Reflection paper on statistical methodology for the comparative

assessment of quality attributes in drug development


Status

• 1st draft in preparation

(EMA WPs consulted)

• start of public consultation Q2/3 2016

• Case studies

• Convergence with other Regulators

(e.g. FDA)

RP on statistical methodology on quality attributes


to reflect when and how far inferential statistical methods can

assist comparative evaluation of quality attributes data

• application of statistical methods comparability, biosimilarity

assessment

• limitations (non-representative nature of retrievable sample data; e.g.

batch number, variability)

• choice of characteristics to be compared

• understanding sources of variability in quality data and 'the unit of

observation'

• Random Sampling / Experimental Approach

• Definition/justification of an equivalence/similarity criterion, acceptance

range

• other…

Process validation Guideline - finalised


http://www.ema.europa.eu/ema/index.jsp?curl=pages/regulation/general/general_content_0003

55.jsp&mid=WC0b01ac0580028e8b

Acknowledgments


Dolca Fabregat-Montfort

Ina-Christine Rondak

Martijn van der Plas

Steffen Gross

Christian Schneider

Pascal Venneugues

Klara Tiitso

Peter Richardson

Thank you for your attention

http://www.ema.europa.eu/ema/

Dr Veronika Jekerle

Quality Office

European Medicines Agency

30 Churchill Place • Canary Wharf • London E14 5EU • United Kingdom

Email [email protected]

Send a question via our website www.ema.europa.eu/contact

Further information

Follow us on @EMA_News

http://www.ema.europa.eu/ema/

mailto:[email protected]

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