biosimilars, biobetters and beyond: coping with an ... · biosimilars, biobetters and beyond:...

12/18/2011

2011 IETF Day 3 Skilton 1

Biosimilars, Biobetters and Beyond:

Coping with an Accelerating

Biotherapeutic Market

©2010 Waters Corporation | COMPANY CONFIDENTIAL

St John Skilton, Ph.D., Ying Qing Yu Ph.D., Weibin Chen Ph.D.

Waters Biopharmaceutical SciencesRakesh KS, Deepalakshmi PD - Waters India Pvt Ltd

India Executive Technology ForumMilford, December 2011

©2010 Waters Corporation | COMPANY CONFIDENTIAL

12/18/2011


Terminology

‘Bio-betters’

‘Bio-generics’

‘subsequent-entry biologicals/biologics’

‘me-too biologicals/biologics’

‘similar biopharmaceuticals’

©2011 Waters Corporation | COMPANY CONFIDENTIAL 3

‘Follow-on Biologics’‘2nd Generation mAbs’

‘noninnovator proteins’

Terminology

‘Bio-betters’

‘Bio-generics’

‘subsequent-entry biologicals/biologics’

‘me-too biologicals/biologics’

‘similar biopharmaceuticals’

BIOSIMILARS


‘Follow-on Biologics’‘2nd Generation mAbs’

‘noninnovator proteins’

12/18/2011


Overview

Market Factors

Regulatory Positions

Deploying

Factors


Deploying New Tools

Return on Investment

Overview

Market Factors


Deploying

Factors


Deploying New Tools


12/18/2011


Top Prescription Drug Sales: Proteins Increasing

Biomols 20081 of top 5 1 of top 5 5 of top 10

28% of top 100

©2011 Waters Corporation | COMPANY CONFIDENTIAL 7 EvaluatePharma.com

Top Prescription Drug Sales: Proteins Increasing

Biomols 20081 of top 5 1 of top 5 5 of top 10

28% of top 100


Biomols 20145 of top 5 7 of top 10

50% of top 100

EvaluatePharma.com

12/18/2011


Antibodies represent the next biotherapeutic class open to European biosimilar competition (McKinsey Report)


Anti-infectivePEGIntron

Erbitux

Pegasys

5.7AvastinRituxanHerceptin Remicade

mAbs are a major class of biotherapeutic soon open to US biosimilar competition

2009 sales ($Billion)

Oncology

Endocrine

Rheumatology

Erbitux

NovoRapid/NovoLog

4.3Lantus Neulasta

Avonex Rebif

5.7

6.1Enbrel

Humira

Levemir

Norditropin SimpleXx

NovoMix

Humalog1.1

0.8


Blood

CNS

Modified slide from McKinsey and CompanyData Source: Evaluate Pharma

201520142013201220112010

Rebif

NovoSeven KogenateProcrit/Eprex

Tysabri

20202019201820172016

Monoclonal AbOther protein

US Patent Expiration

2021

12/18/2011


World-wide Sale of Biologics Coming Off Patent

20

Significant businessopportunity for

biosimilars

58($B)

17

9


2009 2010 20132011 2012 2014 2015 Total

1 4 2 3 9 167 44# ofmlc

1 2 1

8

From: Patricia Seymour, The Challenges of Demonstrating Comparability of Biosimilar Products

Pharmaceutical Growth Biopharmaceuticals vs. Small molecules vs. Biosimilars

Projected Biosimilar

2015

$3.7 Bn

Biosimilar Market

2010


http://pharmtech.findpharma.com/pharmtech/News/Biosimilars-Market-will-be-Worth-37-Billion-by-201/ArticleStandard/Article/detail/713914?contextCategoryId=35097

$234 M

12/18/2011


Cost Estimates High …But Profitability High

Development Cost estimates:

75 250 Milli USD


— 75 - 250 Million USD

— 7 – 8 years

Highly profitable outcomes

Overview

Market Factors


Deploying

Factors


Deploying New Tools


12/18/2011


Worldwide Legislation

“Although copy versions of original biopharmaceuticals are

already available in different parts of the world, there are no

consistent worldwide requirements for their registration”.

o (Yet)


http://www.nature.com/nbt/journal/v29/n8/full/nbt.1936.htmlNature Biotechnology; 29, 690–693, 2011. doi:10.1038/nbt.1936

Worldwide Harmonisation is Getting Closer

“Steps toward a global,

harmonized regulatory approach

may already be in sight, with the

World Health Organization (WHO)

scheduled to finalize its guidelines

on the evaluation of similar

biotherapeutic products in 2011.

According to EGA, the guidelines

ill b f d d h


will be founded on the same

basic scientific principles as in

the EU”. Stephanie Sutton, ‘Call for "Workable" Guideline

for Biosimilar mAbs’; September 9, 2010.

PharmTech.com.

12/18/2011


Current EU Legislation is ‘Clear’

EU: biosimilar is a copy version of the reference product

“D t t d i il it i h i h i l h t i ti “Demonstrated similarity in physicochemical characteristics, efficacy and safety, based on a comprehensive comparability exercise”

Acceptance of variability ('microheterogeneity').


EMA – Established Biosimilar Pathway since 2005

Established pathway allows biosimilars with limited Clinical Trials

2009 workshop debated what ‘similarity’ meant

— Agreed with previous model of ‘well-characterised b h d ’


biotherapeutic product’

— Confirmed that ‘comparability’ concept was appropriate

12/18/2011


Regulator Readiness – FDA is Experienced but Only Now Devising guidelines

BPCI act of 2009 established a framework to develop guidelines

FDA outlined a ‘case-by-case’ principle - no formula exists for biosimilars:

— ‘it’s unlikely that a “one size fits all” systematic assessment of biosimilarity can be

developed’

— ‘ a “totality of the evidence” approach by evaluating more attributes and


— …a totality of the evidence approach, … by evaluating more attributes and

combinations of attributes at greater sensitivities with multiple

complementary methods.

— ‘… the scope and extent of [clinical] studies may be reduced further if more

extensive fingerprint-like characterization is used’.

— ‘Immunogenicity remains a critical factor when assessing biosimilarity’

A Real Biotech Story – Genzyme

Change of reactor led to Change in Glycosylation Pattern

4000L

160L

Reactor

Glycosylation Pattern BGlycosylation

Pattern A

160 L Reactor


Genzyme receives PDUFA date from FDA for its Biologics License Application for Lumizyme - 21. January 2010 23:52 http://www.news-medical.net/news/20100121/Genzyme-receives-PDUFA-date-from-FDA-for-its-Biologics-License-Application-for-Lumizyme.aspx

Reactor

4000L Reactor

12/18/2011


A Real Biotech Story – Genzyme (now Sanofi)

Biotech company had to re-submit as a new product

Glycosylation B Not 4000L 160 L R t Glycosylation B Not

COMPARABLE to Glycosylation A

4000L React

or

Glycosylation Pattern B

Glycosylation Pattern A

Reactor


Genzyme receives PDUFA date from FDA for its Biologics License Application for Lumizyme - 21. January 2010 23:52 http://www.news-medical.net/news/20100121/Genzyme-receives-PDUFA-date-from-FDA-for-its-Biologics-License-Application-for-Lumizyme.aspx

US: New Clinical Trials

US: Resubmission for New Product

EU: (fast) Submission as Biosimilar

EU: Limited Clinical Trials

ESTIMATED COST > 40 M USD

Overview of Types of Biotherapeutic

Size and Complexity increase means that analytical tools need to be able to cope

mAb

150kDaEPO

Conjugated mAb

150kDa+

Blood Factors

250kDa


Insulin

6kDa

GCSF

19kDa

34kDa

PEG-Interferon

20kDa

150kDa+

12/18/2011


Overview

Market Factors


Deploying

Factors


Deploying New Tools


Biosimilar Screening Workflow

Intact mAb LC/MSE

ReductionAlkylationDigestion

VariantProfiling

Intact mAb LC/MS

LC/MSE

Peptide Map

Reduction

Digestion

PNGaseF DeglycosylationHILIC Glycan SPE2AB Label & CleanupHILIC-FLR (MS?)

Higher OrderStructure

Host CellProteins

Stability/Formulation


Released Glycan (FL)LC & HC

LC/MS

Bioanalysis

QCTesting

12/18/2011


Overview

Market Factors


Deploying

Factors

‘Profile’: EPO Glycan profiling

Similarity: Sequence Variants


Deploying New Tools


Immunogenicity: Host Cell Proteins

Comparability in Higher Order

Structure

Overview

Market Factors


Deploying

Factors




Deploying New Tools




Structure

12/18/2011


Critical Tool: UPLC/FL/(MS) methodology -First UPLC 2AB method to Market, Best in Class

Extensive work on complex glycoproteins from Waters and International collaborators

2011 poster


http://www.waters.com/webassets/cms/library/docs/720004019en.pdf

2011 apps note

Waters India EPO Biosimilars Comparison Poster – UPLC/ FL/ MS

EPO poster at NCBS, Bangalore, India, 2nd International Symposium on Mass Spectrometry in Life Sciences


http://www.ncbs.res.in/node/497

12/18/2011


Waters India: Glycan analysis of three biosimilars of EPO using UPLC FLR MS

Biosimilar 1

Biosimilar 2

Biosimilar 3


High reproducibility of UPLC/ FL/ MS allows:— Accurate determination even at low abundance

— Relative quantities in a complex profile

Greater security of information in a challenging marketplace

Overview

Market Factors


Deploying

Factors




Deploying New Tools




Structure

12/18/2011


Characterisation Workflow as Viewed in 2006 (Barnes et al, Lilly)

No Automated processing existed in 2006existed in 2006— Proteomics tools

predominated

MS/MS was at that time not routinely used in the industry for peptide


Barnes et al, 2006; Fig 2.

mapping

Characterisation Workflow Improvements in the Modern Era – Waters 2008 - 2011

Major Workflow advantages with modern UPLC/MS

Deconvolution Required

Optimised, automated i ith M E t

Protein Automated Intact mass

Denaturation, Reduction, Alkylation


processing with MaxEnt

Provision of Intact Mass kit

12/18/2011


2009 Studies on Marketed Biotherapeutics with BPLx and Intact Mass Kit


http://www.waters.com/webassets/cms/library/docs/720

002911en.pdf

HC

LC

HCSpectrum

LCSpectrum

Natalizumab, IgG4

Method Development Simplicity: GenericSEC/UV/MS Method for Reduced IgGs

ACQUITY UPLC™ BEH200 SEC 1.7 µm

4.6x300 mm Column, p/n: 186005226

IgG2

Adalimumab

Bevacizumab

Infliximab

TIC


HC-HC

Rituximab

PTG1

Trastuzumab 10 min

ASMS2011 Poster

12/18/2011





Abandoned?




How to QUANTIFY?




Efficiency NeededAbandoned?


Enzymatic Digestion


One Analysis for


UPLC/ MSE

Automated Disulfide Bond

Mapping

Automated PTM Identification and

Quantification

ymultiple results

Automated Sequence Validation

12/18/2011


Critical Tools: BiopharmaLynx™ and UNIFI™ - First to Market, Best in Class

Purpose-built

Built in collaboration with our partners Built in collaboration with our partners

Ongoing development


Automated EPO comparisons: Protein sequence coverage between Biosimilar 1 and 2

Automated Processing of UPLC/ MSE data

— Early Adopter quote: ‘This has saved us eight weeks of data analysis per sample’

— Performs multiple automated tasks (Intact Mass, Peptide Mapping, Disulfide Bond Mapping)

Colour-coding used to indicate whether ‘control’ and ‘analyte’ have the same match

C t


Line indicates fragment ion data available to validate sequence

Coverage measurement

12/18/2011


UPLC/ MSE: Automated Peptide map Processing of three Biosimilars of EPO

Manual processing would be prohibitively time-consuming

BPLx automates the process for peptide maps— Provides validation of peptide sequence

Biosimilar 1 Sequence Coverage: 96.4%


Provides validation of peptide sequence



Spectrum comparison between Biosimilar 1 and 2-difference plot

Automated, Rapid Visualisation of extent and

location of differences – using objective criteria

justifiable to the regulatorsj g


12/18/2011


Overview

Market Factors


Deploying

Factors




Deploying New Tools




Structure

Deconvoluted Spectra of the Heavy Chain

INNOVATOR

For each pair of forms the mass difference is consistent— Each (glyco)form of the

Biosimilar sample is approx. 32Da less than the corresponding form in th C t l


the Control

The pairs appear to match up for all variants.

32Da 32Da 32DaBIOSIMILAR

12/18/2011


LCMSE Peptide Map Comparison

Innovator mAb

Result: No obvious, gross differences between antibody maps


“Biosimilar” mAb

MIRROR MAP shows Difference Due to Unknown Peak in Biosimilar Sample

Changes clear in mirror plot INNOVATOR

(chromatogram)

Significant Differences are visually salient

BIOSIMILAR(chromatogram)

INNOVATOR(spectrum)


New, Unknown Peak in BIOSIMILAR sample at

m/z 1872.96

Peptide T34-35 (missed cleavage) detected in INNOVATOR sample not

detected in BIOSIMILAR sample

BIOSIMILAR(spectrum)

12/18/2011


Heavy Chain Sequence Coverage –INNOVATOR mAb Sequence T35: EEMTK


Coverage Map for Heavy Chain for Innovator sample is high and mAb Sequence confirmed

The T35 peptide is correctly identified and sequence confirmed in the Innovator product – but not in the Biosimilar

MSE Spectra of INNOVATOR mAb T35 (EEMTK)

BiopharmaLynx automatically provides sequence confirmation thanks to UPLC/ MSE data/


T35 (EEMTK), RT = 3.86 min, Innovator Sample

12/18/2011


Heavy Chain Sequence – Biosimilar Sample with Drugbank mAb Sequence

Sequence information for the Innovator Sample was copied

from a peer-reviewed paper published after submission of IPfrom a peer reviewed paper published after submission of IP

HT35 sequence is confirmed by MSE



Sequence information for the Innovator Sample was copied

from a peer-reviewed paper published after submission of IPfrom a peer reviewed paper published after submission of IP

HT35 sequence is validated

Primary AA sequence in Drugbank for this mAb indicated that

there was a difference at that position

HT35 sequence in Drugbank is –DELTK-


The same dataset was then re-analysed with the

alternative sequence

UPLC/ MSE collected information on ALL ions

12/18/2011



Coverage Map for Heavy Chain for Biosimilar sample using Drugbank Sequence


HT35 peptide now correctly identified

No further sample acquisition was necessary as the MSE

data was already acquired – re-processing with the alternative sequence was completed in minutes.

Mirror View of T35s in BiopharmaLynx Processed data

T35 (EEMTK)Innovator SampleInnovator Sample


T35 (DELTK)‘Biosimilar’ SamplePeptides in each sample can

now be correctly attributed to their respective peptide

sequences purely by reprocessing the data with

new information

12/18/2011


Sample Handling is Critical to Accurate Results

mAbs 2:4, 1-16; July/August 2010; © 2010 Landes Bioscience


Overview

Market Factors


Deploying

Factors




Deploying New Tools




Structure

12/18/2011


Comparability and Biosimilars: BMS and Biogen Idec use of HDX

2011


2011 poster

Biogen Idec Paper on the Advantages of Adopting HDX for Comparability (2011)

Biogen Idec early Adopters of Waters HDX

Fig 6 from paper example of rapid and automated processing of UPLC/ MSE data for a Totality of information not available from traditional MS techniques— LOCALISATION of differences and measure of AMOUNT of difference

simultaneously

The Utility of Hydrogen/Deuterium Exchange Mass Spectrometry

i Bi h ti l C bilit


in Biopharmaceutical Comparability Studies. DAMIAN HOUDE, STEVEN A.

BERKOWITZ, JOHN R. ENGEN. JOURNAL OF PHARMACEUTICAL SCIENCES, VOL.

100, NO. 6, MAY 2011http://onlinelibrary.wiley.com/doi/10.10

02/jps.22432/abstract

12/18/2011


Comparability and Biosimilars: BMS use of HDX for PEGylated GCSF

Application of Waters ‘Butterfly Plot’ to a real-Butterfly Plot to a realworld situation

— Automation of data processing

— MONTHS of time savings over manual workflow

— Objective analysis of


Objective analysis of ‘difference’ vs (subjective) human interpretation

Higher Order Structure and Protein Analytics – Regulatory Authorities

“Our current ability to predict the potency of biologics would be Our current ability to predict the potency of biologics would be

enhanced if we had improved ability to measure and

quantify the correct (major) three-dimensional structure,

aberrant three-dimensional structures (misfolding), and the

distribution of different three-dimensional structures”.


• [TESTIMONY BEFORE THE SUBCOMMITTEE ON TECHNOLOGY AND INNOVATION COMMITTEE ON

SCIENCE AND TECHNOLOGY U.S. HOUSE OF REPRESENTATIVES. SEPTEMBER 24, 2009]

Steven Kozlowski, M.D.Director, Office of Biotechnology Products, Office of Pharmaceutical Science. CDER

12/18/2011


Basics of BiopharmaLynx™ Display for Automated Disulfide Bond Mapping

Each Peak labeled with Peptide Number


Hoverbox for Peptide Information

‘=‘ Symbol represents S-S bond

BSA - Automated assignment of Expected Peptides from Tryptic Digest

With BiopharmaLynx™ All 9 peptides with expected S-S bonds are assigned automatically shown by ‘Peptide1=Peptide2’

‘h b ’ h d k

T07=T07

T08=T10=T12

T14=T21=T22T39=T41=T42

T63=T66=T67

‘hoverboxes’ show sequences at processed peak apex


T28=T37 T44=T50=T51 T54=T61=T62

T69=T76=T77

12/18/2011


Overview

Market Factors


Deploying

Factors




Deploying New Tools




Structure

Host Cell Protein Work - 2011


http://www.waters.com/webassets/cms/library/docs/720004043en.pdf

http://www.waters.com/webassets/cms/library/docs/thp637.pdf

12/18/2011


Waters HCP analysis with LC/MSWorkflow Overview - Discovery

Protein mixture is digested (Trypsin) to identify the peptides by LC/MS

Complexity of mixture is reduced by using online two-dimensional chromatographychromatography

Uses established Informatics tools (PLGS) to:

— Identify the individual proteins by matching [tryptic] peptides to database entries

— Calculate protein concentration based on signal response

— Display and rank matches in a protein-centric format


Perform online 2D LC/MSE to discover impurity proteins.

Spike mixture with known proteins and enzymatically digest

Identify proteins and calculate concentrations

Key Figure for HCP study

Section of table from HCP study

— Multiple cell lines Specific Proteins are identified

Prot Proteinno Description I II III IV

1 Nascent polypeptide associated complex subunits Mus musculus 50 17682 Nucleolin Mesocricetus auratus 2613 Heterogeneous nuclear ribonucleoprotein isoforms Mus musculus 12274 Elongation factor isoforms Mus musculus 142 665 20495 Procollagen C endopeptidase enhancer 1 Mus musculus

CHO-S Cells

— Multiple purification schemes

— Null cell lines

are identified

Protein concentration is measured


5 Procollagen C endopeptidase enhancer 1 Mus musculus6 Laminin subunits Mus musculus7 Actin isoforms Mesocricetus auratus 287 1298 Clusterin Mus musculus 537 2329 Nidogen 1 Mus musculus

10 Glyceraldehyde 3 phosphate dehydrogenase Mes auratus 153 59 7511 Splicing factor proline and glutamine rich Mus musculus 19812 Glycogen phosphorylase b rabbit - PHO (800 fmoles) 778 778 778 77813 Plasminogen activator inhibitor 1 RNA binding protein Mus musculus 226

Spiked standard with measured recovery

Consistentmeasurement across

all conditions

12/18/2011


High Throughput MRM Monitoring and absolute quantitation

Absolute Quantification with stable isotopically

labelled peptides as internal standards

— Targeted for specific proteins

— Simple transfer of MSE conditions to MRM

methods

Provision for alternative peptides to be chosen

Assay extremely simple to update/ change in

minutes


minutes

MRM Monitoring of an HCP level in MRM Monitoring of an HCP level in 10 PTG1 mAb Batches from CHO10 PTG1 mAb Batches from CHO

mAb Batch (in Triplicate)

1 2 3 4 5 6

7 8

9 10


12/18/2011


Overview

Market Factors


Deploying

Factors


Deploying New Tools


Engineering Routine High Performance Mass Spectrometry

Automating tuning and calibration Integrated fluidics system


Automated LC/MS system checks

Continuous background monitoring

Updated electronics for diagnostics

No tools needed

12/18/2011


Analytical Platforms across Analytical Platforms across Biotherapeutic OrganizationsBiotherapeutic Organizations

MassLynx Usage

Empower Usage

Discovery

Development

Discovery

Development

MassLynx Usage

Few compliance issues GxP compliance Regulatory Compliance


Production

Post-Approval

QC/QA

Production

Post-Approval

QC/QA

VALIDATIONSTABILITYCHARACTERIZATION

Analytical Platforms across Analytical Platforms across Biotherapeutic OrganizationsBiotherapeutic Organizations

Empower Usage

Discovery

Development

Discovery

Development

Few compliance issues GxP compliance Regulatory Compliance


Production

Post-Approval

QC/QA

Production

Post-Approval

QC/QA

VALIDATIONSTABILITYCHARACTERIZATION

12/18/2011


Biopharmaceutical System Solution

LC and LC/MSE Peptide Mapping


LC/MS Intact Mass

RP, SEC and IEX

Your Success is Our Mission

Satisfy the Regulator

Raise the bar against the Competition

Increase safety for patients


SECURE YOUR POSITION IN THE MARKET

12/18/2011



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