SEPTEMBER 17TH, 2015

A QUALITY DRIVEN BIOSIMILAR DEVELOPMENT

AN AVERSE RISK APPROACH

Amman, Jordan

September 2015 /

AFTER LUNCH TEST: FIND THE DIFFERENCE

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?

September 2015 /

CHALLENGES IN BIOSIMILAR DEVELOPMENT

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Structure

Biological Activity

Stability

Safety

Efficacy

Originator Similarity of Biosimilar

September 2015 /

OUTLINE

Understand the Complexity of Biologics

Define the Originator Quality Target Product Profile

Achieve Biosimilarity

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September 2015 /

N-GLYCOSYLATION BIOSYNTHESIS

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September 2015 /

COMPLEXITY OF N-GLYCOSYLATION IN HUMANS

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September 2015 /

TYPICAL BIOSIMILAR DEVELOPMENTS

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Reference Product

Produced in Glycans per Chain

Active Ingredient

Avastin® CHO 1xN (IgG1) Bevacizumab

Avonex® E.coli Interferon beta-1a

Benlysta® NS0 1xN (IgG1) Belimumab

Betaseron® E.coli Interferon beta-1b Copaxone chemical Glatiramer acetate

Enbrel® CHO;

3xN, 10xO (IgG1-Fus.)

Etanercept

Epogen®, Eprex® CHO; 3xN, 1xO Erythropoietin

Erbitux® SP2/0; 2xN (IgG1) Cetuximab

Gonal F® CHO; 4xN

Follicle-stimulating hormone

Herceptin® CHO; 1xN (IgG1) Trastuzumab

Humira® CHO, 1xN (IgG1) Adalimumab

Lantus® E.coli Insulin

Lucentis® E.coli Ranibizumab

Neupogen® E.coli Filgrastim

Reference Product Produced in Glycans per Chain

Active Ingredient

Nutropin® E.coli Somatropin

Octocog alpha® CHO 25xN+12xO Factor VIII

Pegasys® E.coli PEG-Interferon alpha-2a

Proleukin® E.coli Interleukin-2

Prolia® CHO 1xN (IgG2) Denosumab

Remicade® SP2/0 1xN (IgG1) Infliximab

Rituxan® CHO 1xN (IgG1) Rituximab

RoActemra® CHO 1xN (IgG1) Tocilizumab

Roferon® E.coli Interferon alpha-2a

Simponi® SP2/0 1xN (IgG1) Golimumab

Soliris® SP2/0 1xN (IgG2/4) Eculizumab

Stelara® SP2/0 1xN (IgG1) Ustekinumab

Tysabri® NS0 1xN (IgG4) Natalizumab

Xolair® CHO 1xN (IgG1) Omalizumab

Yervoy® CHO 1xN (IgG1) Ipilimumab

September 2015 /

INLFUENCE DURING MANUFACTURING

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Glycosylation

Aggregation

Deamidation

HCPs

K-Clipping

Oxidation

Fragmentation Disulfide shuffling

Glycation

Degradation

Clone development

September 2015 /

QUALITY ATTRIBUTES OF MABS

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September 2015 /

ANALYTICAL ASSAYS FOR BIOSIMILARITY ANALYSIS

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Structure/ Sequence • N- and C-terminus • Amino acid analysis • Peptide Mapping and Sequencing • Disulfide Linkage Analysis • Intact mass (reduced and middle down)

Aggregation • SEC • AUC • SDS-PAGE • AF4

Glyosylation • Monosaccharide Analysis • Oligosaccharide Analysis • Intact Mass • MALDI-TOF • HILIC-HPLC/MS

Higher Order • CD • FT-IR • NMR • H/D Exchange • X-Ray

Surface Charge • IEX • CZE • IEF/c-IEF

Activity • Bioassays • Binding assays

September 2015 /

OVERVIEW GLYCOSYLATION ANALYSIS

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Chromatographic Profiles

Saccharide Content

Glycan Pool Analysis

GlycosylationSite/ Site Occupancy/

Site Specific Pool

Glycan Structure

w/wo Deglycosylation w/wo Desialisation Degylcosylation w/wo Deglycosylation

Site specific protein fragment Exoglycosidase Defucosylation

IEF gel/cIEF Monosaccharide

Content by HPAEC-PAD

HILIC-HPLC HPAEC-PAD

Glycosylation Site and Site Occupancy by Peptide Mapping LC-ESI-MS/MS,

LC-UV

Linkage Analysis GC-MS

IEX-HPLC/RP-HPLC Sialic Acid Content by HPAEC-PAD

MALDI-MS of Released Glycans

Glycan distribution per site by Peptide Mapping

LC-ESI-MS/MS

LC-ESI/MALDI-MS/MS

2D PAGE 1D PAGE/cGE

Intact molecular weight by MALDI-MS

or LC-ESI-MS

Intact molecular weight by MALDI-MS or LC-ESI-MS NMR

September 2015 /

CHOICE TO SEE BIOSIMILARITY

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Initially fast

High risk

Some Quality Driven Development

Upfront loaded, quality driven

Lower risk

Full Quality Driven Development

Establishing Biosimilarity by scientific principles

Comparability to make a

September 2015 /

BIOSIMILAR DEVELOPEMENT - DEFINING QTPP AND CQA

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Formulation

Upstream Proc Dev

Cell Line Dev

Downstrem Proc Dev

Structural characterization e.g Amino acid sequence and compos. N/C terminus Post translational modification Glycosylation Disulfide linkage Higher order structure Aggregates

Bioassays

Physicochemical properties e.g. Molecular weight or size Isoform pattern Extinction coefficient Electrophoretic pattern Liquid chromatographic pattern Spectroscopic profiles

Ana

lytic

s

Detailed Characterization of Originator Batches

Quality Target Poduct Profile QTPP

May change during Development (e.g. Process Changes)

Critical Quality Attributes Linking Quality to Safety and Effiicacy

Enabling QbD Development Com

para

bilit

y

September 2015 /

MODULES ON REFERENCE MEDICINAL PRODUCT

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QTPP by Originator Monitoring Program

Originator AA verification

Originator Monitoring Program

Media Optimization

Upstream/ Downstream Develop

Eng. Runs and GMP Production

Pre+Clinical Phase I+III Approval Cell Line

Develop

Originator Sourcing and Logistics

Method Qualification

September 2015 /

QUALITY TARGET PRODUCT PROFILE OF 6 RMP

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Originator Batch ABC

Quality Target by Originator Batch EDF

Biosimilar Candidate

Considered not highly similar

Originator Batch ABC

Originator Batch EDF

Biosimilar Candidate

High number/high variability of characterized originator batches increases quality target range for the Biosimilar Candidate

Assumed to be highly similar

Qua

lity

Attr

ibut

e in

%

Qua

lity

Attr

ibut

e in

%

September 2015 /

CHANGE OF GLYCOSYLATION FOR MARKETED PRODUCTS

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Schiestl et al., Nature Biotechnology, Vol 29, No 4 April 2011

Rituxan/Mabthera Enbrel

September 2015 /

ONGOING ORIGINATOR MONITORING

Detailed Monitoring program of e.g. 30 RMP Lots: 10 US, 10 EU, 10 ROW

Sourced internationally with highest variability in Lots and Exp. Dates; to include different Production sites and Process Changes of RMP

Sourced and analyzed in campaigns

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September 2015 /

METHOD QUALIFICATION FOR BIOSIMILARITY

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The assays needs to be suitable to detect small differences

Generation of Stress samples

Testing of Stress samples

Final method qualification with

tested stress samples

As an example to be able to qualify selected assays a variety of stress samples will be generated and tested with two different orthogonal method.

Subsequently the samples will be used as a reference Std. for Method Qualification

September 2015 /

QTPP driven

MODULES ON BIOSIMILAR CANDIDATE

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Pool selection

Clone selection

Media Selection

Process development

Media Optimization

Upstream/ Downstream Develop

Eng. Runs and GMP Production

Pre+Clinical Phase I+III Approval Cell Line

Develop

Standardized and validated release testing

Stability testing

Comparability studies

September 2015 /

QUALITY DRIVEN DEVELOPMENT

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QTPP driven Pool selection

Clone selection

Media Selection

Process development

Initially fast

High risk

Upfront loaded, quality driven

Lower risk

Not an overall cost argument !

September 2015 /

EXERPT OF QTPP

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September 2015 /

DEVELOPMENT BY QUALITY ATTRIBUTES

Quality directed development planned in several iterations:

Clone Pool Analysis Clone Selection Media Optimization

Starting with >100 Samples and selected Quality Attributes

Screening Assays optimized for parallelization, speed and sensitivity

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Targeted attribute

Limits for this attribute

Maybe highly productive pool but not suitable

Clone Pool Single Clones Media Optimization

September 2015 /

EXAMPLE: ANALYSIS FOR CLONE SELECTION

Application: Development of an IgG1 biosimilar • MALDI-MS and subsequent comparison of glycosylation pattern • 96 samples in less than 6h (including ProteinA, Glycan Release, MALDI,

Data Evaluation)

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1605

.6

0.0

0.5

1.0

1.5

2.0

5x10

Inte

ns. [

a.u.

]

1300 1400 1500 1600 1700 1800 1900 2000 2100m/z

G0

High Mannose

G0 F

G1

G1 F

G2

G2 F

Calculated Ratios:G1/G0 and G2/G0 fucosylated/non-fucosylated

September 2015 /

NOT MEETING GLYCOSYLATION FOR BIOSIMILARITY

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-

For biosimilarity analysis, single assays can question biosimilarity.

However, demonstrating biosimilarity needs extensive characterization.

Sample E IA IB IIA IIB IIIA IIIB IV V VII VIII E

Schellekens H et al. Eur J Hosp Pharm Pract 2004;3:43–7

Eprex and Copy Biologics

September 2015 /

WHY QUALITY?

AIM: Mimimize risk starting at early development to

meet biosimilarity in the end • Saves time! • Reduces clinical trials • Mitigates later adaption of process design • Meet the authorities´ expectations

Build close collaboration between partners for optimal results

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September 2015 /

THE BIOSIMILAR GROUP CONCEPT

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June 15-18 2015 /

www.TheBiosimilarsGroup.com

The Biosimilars Group GmbH, Harrlachweg 6, 68163 Mannheim, Germany

Phone: +49 (0) 621 76449064 Email: Ulrike.Konrad@TheBiosimilarsGroup.com

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June 15-18 2015 /

Patent Situation

Development Plan

Market Access

DECISION MAKING

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