Comparability Studies:The Key to a Biosimilar's Success 

Raymond Kaiser, PhD

Global Vice President

BioPharmaceutical CMC Solutions

Covance

2

Presentation Outline

What is “Biosimilarity”

Global Regulatory CMC Expectations

Comparability Expectations

Analytical Approaches to Comparability

“Real World” Observations

Q&A

33

What is a Biosimilar?

Biosimilars are follow-on biologics New versions of innovator

biopharmaceutical products,

following patent expiry

The very nature of a biologic means It is practically impossible for two different

manufacturers to produce two identical biopharmaceuticals if identical host expression systems, processes and equivalent technologies are not used

This has to be demonstrated in an extensive comparability program

4

Biosimilar or Biosimilarity means:

The biological product is highly similar

to the reference product notwithstanding minor differences in clinically inactive components

There are no clinically meaningful differences

between the biological product and the reference product in terms of the safety, purity, and potency of the product

5

Global Regulatory LandscapeEMA (The European Medicines Agency)

Established EMA approval pathway: 14 biosimilars approved* within the product classes of:

human growth hormone granulocyte colony-stimulating factor (G-CSF) erythropoietin

US FDA Biologics Price and Innovation Act passed in 2010

Empowers FDA to develop standards to evaluate and approve biosimilarsFDA February 2012 draft guidance documents

Risk-based “totality-of-the-evidence” approach

Health CanadaGuidance came out in 2010

JapanGuideline came out in 2009

WHOGuideline document came out in 2009

ROWArgentina, Australia (amending), Brazil, Jordan, Korea, Malaysia, Saudi Arabia, Singapore, Taiwan,

Turkey, Venezuela – Guidelines availableColombia, Cuba, India, Mexico, South Africa, and Thailand – Draft Guidelines available

Biosimilar Regulatory Guidance documents and discussions are on-going and expanding globally.

6

Regulatory Environment - EU

Established EMA approval pathway 14 biosimilars approved within the product classes of:

Human growth hormone

Granulocyte colony-stimulating factor (G-CSF)

Erythropoietin

Key EMA guidance documents “Overarching” guideline on similar biological medicinal products

From 2005, currently being updated

Guideline on similar biological medicinal products containing biotechnology-derived proteins as active substances: quality issues

From 2005

Guideline on similar biological medicinal products containing biotechnology-derived proteins as active substances: nonclinical and clinical issues

From 2005, currently being updated to incorporate risk-related approaches

7

Regulatory Environment – US

FDA approval pathway established

Biologics Price and Innovation Act passed in 2010 empowers FDA to develop standards to evaluate and approve biosimilars

FDA February 2012 draft guidance documents Scientific Considerations in Demonstrating Biosimilarity to a Reference Product

- Risk-based “totality-of-the-evidence” approach

Quality Considerations in Demonstrating Biosimilarity to a Reference Protein Product- Overview of analytical factors to consider, includes analytical, physico-

chemical and biological characterization

Biosimilars: Questions and Answers Regarding Implementation of the Biologics Price Competition and Innovation Act of 2009 - Provides answers to common questions that may arise in the early stages

of product development

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General Requirements of FDA Guidance

A 351(k) application must include information demonstrating biosimilarity based on data derived from:

Analytical studies demonstrating that the biological product is “highly similar” to the reference product notwithstanding minor differences in clinically inactive components

Animal studies (including the assessment of toxicity) A clinical study or studies (including the assessment of

immunogenicity and pharmacokinetics (PK) or pharmacodynamics (PD)) that are sufficient to demonstrate safety, purity, and potency in 1 or more appropriate conditions of use for which the reference product is licensed

FDA may determine, at its discretion, that an element described above is unnecessary in a 351(k) application

9

Regulatory Environment - ICH Regions

How do the requirements compare?

US, EU and Japan all require comparability with reference product

Reference product must be registered under the Regulatory jurisdiction In EU this is law FDA will consider non-US registered reference product with studies to bridge to US reference Japan requires Japanese registered product

Early engagement with Regulatory Authority is vital

Extrapolation of indications will be considered

At least one Phase III comparable efficacy study (usually with equivalence design) is required for licence

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Regulatory Comparability PackageExtensive Analytical & In Vitro

Extensive analytical characterization versus reference product(s) If mAb

specificity & affinity to epitope potency using CDC, ADCC &/or functional in vitro cell-based assays

Limited in vivo Nonclinical PK/PD studies, Toxicology (SD/MD), and Immunogenicity studies

(requirement for nonhuman primates?)

Limited Clinical Phase I

comparability of PK/PD & Immunogenicity in volunteers or patients at reference dose and regimen for targeted indications

Phase III comparability for safety and efficacy (single pivotal) based on PK/PD,

surrogate endpoints and clinical outcomes

Extensive Post-Approval REMS and pharmacovigilance planning including patient registry(ies) for

assessment of incidence of specific safety issues

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Development Challenges

Regulatory, manufacturing and marketing complexities

Biosimilar must be highly similar to innovator biologic, which can be difficult to demonstrate as all data for innovator will be lacking

A comparability exercise has to be followed with the innovator product at all levels of product development

Physico-chemical characterization

Biological activity

Preclinical in vivo comparability

Phase I PK and safety

Phase III efficacy and safety

It’s all about comparability

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Biosimilarity?

How close do the proposed biosimilar products (figures B-E) compare to the reference product (figure A)?

Advances in current state-of-the-art analytical methods enhance the likelihood that a product will be highly similar to another product by better targeting the original product's physicochemical and functional properties

‘Biosimilar Biological Product’ Webinar, Rachel Sherman, FDA, 15FEB2012

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‘Biosimilar Biological Product’ Webinar, Rachel Sherman, FDA, 15FEB2012

FDA View

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Biosimilars must be Systematically Engineered to Match the Reference Product

Mark McCamish, Novartis, International Conference on Drug Development, Austin TX, 29FEB2012

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Characterization

Comprehensive Characterization Physicochemical as well as biological

Multiple batches from innovator spanning a number of years

Understand innovator variability Specification changes over life of product

No label change

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Biosimilars at CovanceYear on year doubling of analytical demand for Biosimilars since 2008

Upgraded Protein Chemistry techniques

with additional characterization to demonstrate comparability or understand differences

Moved toward higher defining analytics (e.g. UPLC, LC/MS)

Compounds 1000 500 250 100 20 1

IND/CTA NDA /BLA Approval

DISCOVERY

PreclinicalResearch Phase I Phase II Ph. IIIa IIb Phase IV

DRUG DEVELOPMENT COMMERCIALIZATION

Biosimilar

NBE

Am

ou

nt

of

Ch

ara

cte

ris

atio

n

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Why More Characterization for Biosimilars?

Client has to show high ‘similarity’ to Innovator

“Proving ‘highly similar’ to the reference product often required multiple iterations of process change and physiochemical characterization”

Characterization of the Innovator (beware literature!) Characterization of the Biosimilar Monitor Manufacture

‘Process is the product’ Monitor changes in Innovator

Analytical tools for characterizing biopharmaceuticals and the implications for Biosimilars. S Berkowitz, J Mazzeo, G Jones, Nature review Vol 11, Jul 2012

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Why More Characterization for Biosimilars?

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Protein Heterogeneity Amino Acid Substitution AA Misincorporation (e.g.

METNLE) N- and C-terminal mods Mismatched S-S bonds Folding Truncation Aggregation Multimer Dissociation Denaturation Acetylation Fatty acid acylation Deamidation

Oxidation Carbamylation Carboxylation Formylation -Carboxyglutamylation O-linked Glycosylation N-linked Glycosylation Methylation Phosporylation Sulphation PEGylation etc.

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Analytical Tools to Evaluate Protein Structure (subset)

Protein Functionality Analytical Technique

AA Sequence and Modifications

Mass Spectrometry (MS), peptide mapping, Edman Sequence analysis, chromatographic separations

FoldingS-S bonding, calorimetry, HDX and Ion mobility MS, NMR, CD, FT & Raman spectroscopy, fluorescence, chromatography

Subunit Interactions Chromatography, ion mobility MS

Heterogeneity of size, charge, hydrophobicity

Chromatography, gel & capillary electrophoresis, light scatter

Glycosylation Anion exchange, enzymatic digestion, peptide mapping, CE, MS

PEGylation & isomers Chromatography, peptide mapping

Bioactivity, cellular and animal bioassays

Ligand & receptor binding (ELISA, SPR), signal transduction

AggregationAnalytical ultracentrifugation, size-exclusion chromatography, field flow fractionation, light scatter, microscopy

Proteolysis Electrophoresis, chromatography, MS

Impurities Chromatography, proteomics, immunoassays, PCR

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Mass Spectrometry

Intact mass, comparison of Glycan variants. The difference in 56Da attributed to incorrect amino acid sequence

2222

Hormone Receptor Binding by Biacore

-10

0

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0 50 100 150 200 250 300 350

Time s

Re

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RU

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0 50 100 150 200 250 300 350

Time s

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RU

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0 50 100 150 200 250 300 350

Time s

Re

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iff.

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DS BiosimilarDP Biosimilar

Innovator

Sample Lot No. KD (nM) % Difference1 3012 3993 4121 4152 3503 397

Drug Substance

4.40

Innovator

Biosimilar

Sample Lot No. KD (nM) % Difference1 2022 1723 1181 1382 1883 158

Drug Product

-1.64

Biosimilar

Innovator

2323

Finding the Differences by C-IEF

7.41

9

7.52

4

Mai

n -

7.61

3

7.80

8

7.99

1

8.14

5

Abs

orba

nce

0.00

0.02

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

0.20

0.22

0.24

Minutes

6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40 8.60 8.80 9.00 9.20 9.40

Blue - BiosimilarBlack - Innovator

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Carboxypeptidase Treatment

7.15

9

7.24

9

7.33

3

7.43

0

Mai

n -

7.53

3

7.65

9

7.73

1

Abs

orba

nce

-0.02

0.00

0.02

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

0.20

0.22

0.24

0.26

0.28

Minutes

6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40 8.60 8.80 9.00 9.20 9.40 9.60

Blue - InnovatorBlack - Biosimilar

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Forced Degradation

6.79

8

6.92

76.

996

7.13

8

7.26

5

7.38

2

7.48

1M

ain

- 7.

531

7.60

3

7.70

9

7.80

5

7.90

6

8.04

1

Abs

orba

nce

-0.010

0.000

0.010

0.020

0.030

0.040

0.050

0.060

0.070

0.080

0.090

0.100

0.110

0.120

0.130

Minutes

6.00 6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40 8.60 8.80 9.00 9.20 9.40 9.60 9.80

Black - InnovatorBlue - Biosimilar

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Impact of Glycosylation?

Blue- InnovatorBlack- Biosimilar

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Simplify by De-sialylation?A

bsorb

ance

-0.02

0.00

0.02

0.04

0.06

0.08

0.10

0.12

0.14

0.16

0.18

0.20

0.22

0.24

0.26

0.28

0.30

Minutes

3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50

Blue- InnovatorBlack- Biosimilar

2828

N-Glycan SimilarityE

nerg

y

0.00

200.00

400.00

600.00

800.00

1000.00

1200.00

1400.00

1600.00

1800.00

2000.00

2200.00

Minutes

8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00

G0F

G1F

G1F’

G0 G1G1’

Man5

G2F Blue- InnovatorBlack- Biosimilar

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Mass Spectrometry

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Peptide Map

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NOTE: Originators make ‘biosimilars’ every time there is a manufacturing process change

Aranesp ® (Darbepoetin-alfa)

MabThera®/Rituxan® (Rituximab)

Enbrel ® (Etanercept)

Ref: Schiestl et al. Nat. Biotechnol. 2011; 29: 310-312 (Sandoz)

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Characterization of Commercial Batches of Darbepoetin alfa from the EU

Change in isoform distribution between two sets of batches (expiry date April 2010 and September 2010)

Schiestl et al. Nat. Biotechnol. 2011; 29: 310-312

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Characterization of Commercial Batches Mabthera®/Rituxan® (rituximab)

•Shift in Glycosylation Profile and ADCC Potency

• Differences/shift in glycosylation pattern results in different potency in cell-based assays• Product label remained unchanged –indicating comparable quality

Schiestl et al. Nat. Biotechnol. 2011; 29: 310-312

34

Characterization of Commercial Batches of Enbrel

• Shift in Glycosylation Profile

• Differences/shift in glycosylation pattern• Product label remained unchanged –indicating comparable quality

Schiestl et al. Nat. Biotechnol. 2011; 29: 310-312

3535

Demonstrating Comparability in an Ideal World?

Product heterogeneity is clearly understood, variants are easily isolated and characterized Product variants and related impurities Process related impurities

Smooth manufacturing scale up, no process changes

Methods ready to demonstrate comparability without development. “One size fits all”

Enough time and a ‘crystal ball’ to know what to look for

3636

What does This Look Like in the ‘Real World’?

Biosimilars are “snowflakes”. No two are the same

Incomplete comparability characterization

Experience rapid time pressures

Final formulation often undecided

Manufacturing difficulties with scale up

Fast turn around in-process sample analysis

Rapid development and validation for ‘discriminating’ assays

Need justification and risk assessment for observed differences

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Other ‘Real World’ Observations II

Stress stability studies (w/ multiple time points) needed

Multiple lots of reference product often used. Understand impact of ref. product ‘shelf-life’ on results. Justify use

Reference product isolation procedure can impact results

Comparability continues even after release

GMPs apply for biosimilar product development

Meet with BOHs as early as possible to discuss your analytical control strategy

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Other ‘Real World’ Observations IIIProtein analysis requires an ‘integrated’ set of analytical

methodsEvaluate all domains and protein modifications

Use orthogonal analytical methods to confirm observations and expectations.

State-of-the-art techniques are expected

Realize each analytical method has strengths and weaknesses:Spectral methods measure averages

Qualitative vs. quantitative?

Variable sensitivity

etc.

3939

Summary

Biosimilar regulatory guidelines continue to evolve

Demand for biosimilar CMC development continues to grow

Demonstrating comparability requires extensive analysis pre- and post-clinically. Must completely characterize innovator product

We are making progress linking some, but not all, biological properties to critical quality attributes; therefore, Given a gradation of a biologics complexity, a one size fits all

strategy for biosimilars will not be possible

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