Comparability Studies: The Key to a Biosimilar's Success

Raymond Kaiser, PhD Global Vice President BioPharmaceutical CMC Solutions Covance

Presentation Outline
What is Biosimilarity

Global Regulatory CMC Expectations

Comparability Expectations

Analytical Approaches to comparability

Real World observations

Q&A

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

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

Global Regulatory Landscape

EMA (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 biosimilars Risk-based totality-of-the-evidence approach

FDA

February 2012 draft guidance documents

Health Canada
Guidance

came out in 2010 came out in 2009 document came out in 2009

Japan
Guideline

WHO
Guideline

ROW
Argentina,

Australia (amending), Brazil, Jordan, Korea, Malaysia, Saudi Arabia, Singapore, Taiwan, Turkey, Venezuela Guidelines available Colombia, Cuba, India, Mexico, South Africa, and Thailand Draft Guidelines available

Biosimilar Regulatory Guidance documents and discussions are on-going and expanding globally.
5

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 biotechnologyderived proteins as active substances: quality issues

From 2005

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

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

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, physicochemical 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

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
8

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

Regulatory Comparability Package

Extensive 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 comparability for safety and efficacy (single pivotal) based on PK/PD, surrogate endpoints and clinical outcomes

Phase III

Extensive Post-Approval

REMS and pharmacovigilance planning including patient registry(ies) for assessment of incidence of specific safety issues

10

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

Its all about comparability

11

Biosimilarity?

How close do the proposed biosimilar products (figures BE) compare to the reference product (figure A)? Advances in current state-ofthe-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

12

FDA View

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

13

Biosimilars must be Systematically Engineered to Match the Reference Product

Mark McCamish, Novartis, International Conference on Drug Development, Austin TX, 29FEB2012
14

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

15

Biosimilars at Covance
Year 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
DISCOVER Y

1000

500

250

100

20

DRUG DEVELOPMENT

COMMERCIALIZATIO N

Research

Preclinical

Phase I Phase II

Ph. IIIa IIb

Phase IV

Amount of Characterisation

IND/CTA

NDA /BLA

Approval

Biosimilar
NBE

16

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

17

Why More Characterization for Biosimilars?

18

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.

19

Analytical Tools to Evaluate Protein Structure (subset)

Protein Functionality
AA Sequence and Modifications Folding

Analytical Technique
Mass Spectrometry (MS), peptide mapping, Edman Sequence analysis, chromatographic separations S-S bonding, calorimetry, HDX and Ion mobility MS, NMR, CD, FT & Raman spectroscopy, fluorescence, chromatography Chromatography, ion mobility MS Chromatography, gel & capillary electrophoresis, light scatter Anion exchange, enzymatic digestion, peptide mapping, CE, MS Chromatography, peptide mapping Ligand & receptor binding (ELISA, SPR), signal transduction Analytical ultracentrifugation, size-exclusion chromatography, field flow fractionation, light scatter, microscopy Electrophoresis, chromatography, MS Chromatography, proteomics, immunoassays, PCR

Subunit Interactions
Heterogeneity of size, charge, hydrophobicity Glycosylation PEGylation & isomers Bioactivity, cellular and animal bioassays Aggregation Proteolysis Impurities
20

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

21

Hormone Receptor Binding by Biacore

RU 80 RU 80 70 70 60 60 50 50

Resp. Diff.

Resp. Diff.

40

DS Biosimilar

40

30

30

DP Biosimilar

20

20

10

10

-10 0 50 100 150 Time 200 250 300 350 s

-10 0 50 100 150 Time 200 250 300 350 s

RU 90

Sample
70

Biosimilar

50

Innovator

Resp. Diff.

Innovator
30

Drug Substance Lot No. KD (nM) 1 301 2 399 3 412 1 415 2 350 3 397 Drug Product Lot No. KD (nM) 1 202 2 172 3 118 1 138 2 188 3 158

% Difference

4.40

Sample Biosimilar

% Difference

10

-1.64

Innovator
-10 0 50 100 150 Time 200 250 300 350 s

22

Finding the Differences by C-IEF

0.24 0.22 0.20 0.18 0.16 0.14

Main - 7.613

Absorbance

0.12 0.10 0.08 0.06 0.04 0.02 0.00 6.60 6.80 7.00 7.20 7.40 7.60

7.808

7.991

Blue - Biosimilar Black - Innovator

7.524 7.419

7.80

8.00 Minutes

8.145
8.20

8.40

8.60

8.80

9.00

9.20

9.40

SampleName: SB03iCE12-ADM3011P1 5C Lyo Date Acquired: 15-Dec-2010 20:16:15 -05:00 Result Id: 1581 SampleName: SB03iCE08-WL00039785 5C Lyo Date Acquired: 15-Dec-2010 18:55:04 -05:00 Result Id: 1580

23

Carboxypeptidase Treatment
0.28 0.26 0.24 0.22 0.20 0.18 0.16 0.14 0.12 0.10 0.08 0.06

Absorbance

7.430

Main - 7.533

7.249

0.04 0.02 0.00 -0.02 6.40 6.60 6.80 7.00

7.333

Blue - Innovator Black - Biosimilar

7.659 7.731

7.159

7.20

7.40

7.60

7.80

8.00 Minutes

8.20

8.40

8.60

8.80

9.00

9.20

9.40

9.60

SampleName: SB07 iCE 05-WL00039785 9M RecoP3 Date Acquired: 21-Jul-2011 18:53:27 -05:00 Result Id: 4011 SampleName: SB07 iCE 09-ADM30011P1 9M RecoP1 Date Acquired: 21-Jul-2011 20:34:30 -05:00 Result Id: 4017

24

Forced Degradation
0.130 0.120 0.110 0.100 0.090

Absorbance

7.265

0.080 0.070 0.060

7.138

7.382

Blue - Biosimilar Black - Innovator

7.481 Main - 7.531 7.603 7.709

0.050 0.040 0.030 0.020 0.010 0.000 -0.010 6.00 6.20 6.40 6.60 6.80 7.00

6.927

6.996

6.798

7.805

7.906

7.20

7.40

7.60

7.80 8.00 Minutes

8.041
8.20

8.40

8.60

8.80

9.00

9.20

9.40

9.60

9.80

SampleName: SB09 iCE 12-ADM3001P1 30C Rec Date Acquired: 20-Sep-2011 21:19:46 -05:00 Result Id: 4716 SampleName: SB10 iCE 13-WL00039785 30C Rec Date Acquired: 20-Sep-2011 21:27:34 -05:00 Result Id: 4758

25

Impact of Glycosylation?

Blue- Innovator Black- Biosimilar

26

Simplify by De-sialylation?
0.30 0.28 0.26 0.24 0.22 0.20 0.18 0.16

Absorbance

Blue- Innovator Black- Biosimilar

0.14 0.12 0.10 0.08 0.06 0.04 0.02 0.00 -0.02 3.50 4.00 4.50 5.00 5.50 6.00 6.50 Minutes 7.00 7.50 8.00 8.50 9.00 9.50

27

N-Glycan Similarity
2200.00

G0F

2000.00

1800.00

1600.00

G1F
1400.00

G2F

Blue- Innovator Black- Biosimilar

Energy

1200.00

1000.00

Man5

G1F

800.00

600.00

400.00

G0

G1 G1

200.00

0.00 8.00 10.00 12.00 14.00 16.00 18.00 Minutes 20.00 22.00 24.00 26.00 28.00 30.00

28

Mass Spectrometry

29

Peptide Map

30

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)

31

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

32

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
33

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
34

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

35

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

36

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

Other Real World Observations III

Protein analysis requires an integrated set of analytical methods
Evaluate 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.

38

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

39

Let us know how we can help you!

DISCOVERY RESEARCH PRECLINICAL DEVELOPMENT PHASE I PHASE III COMMERCIALIZATION PHASE IV Efficacy Model Development/Biomarker Development Immunogenicity, PK, TK Immunotoxicity: CDC & ADCC Tissue Cross Reactivity Pharmacokinetics/Toxicity Viral Clearance Process Development Support, Biomanufacturing Support, Biosafety Testing In vivo/In vitro Biopotency Physicochem & Biological Characterization Serum Production Clinical Feasibility Phase I to Phase III Clinical Trials Clinical Development: Target Population Clinical Development: Special Populations Stability and Release Outcomes/PE Studies Central Labs Data: Safety and Genotyping Post-Marketing Commitments Health Economics Assessment Reimbursement

Molecular Development (Program Management & Clinical) Regulatory Strategy, EMA/FDA Documentation Prep & Meeting Attendance, CTA/IND/BLA Support and Submission

40

Acknowledgments

Our many global clients who have challenged us with various Biosimilar products Sian Estdale, Raymond Donninger, Ji Wu and the rest of the Covance team

41

Thank you . Questions?

raymond.kaiser@covance.com

42