Learning from History: Making Biosimilars a Reality - Genetic ...

Learning from History: Making Biosimilars aRealityLearning from History: Making Biosimilars aRealityBroadcast Date: Thursday, November 1, 2012Time: 1:00 pm ET, 10:00 am PTSponsored by

Learning from History: Making Biosimilars aRealityYour ModeratorJohn SterlingEditor-in-ChiefGenetic Engineering & Biotechnology News

Learning from History: Making Biosimilars aRealityMark McCamish, M.D., Ph.D.Global Head of Biopharmaceutical DevelopmentSandoz Biopharmaceuticals

Making Biosimilars a RealityMark McCamish, MD, PhDHead Global Biopharmaceutical DevelopmentSandoz BiopharmaceuticalsGenetic Engineering & Biotechnology News WebinarNovember 2012a Novartis company

OverviewWhy biosimilars?QbD and the scientific approach to biosimilar developmentClinical trial requirementsSuccesful commercialization broadens patient access5 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

Growing demand drives costs… and threatens tolimit patient access (US example)Estimated daily treatment costs 1in USD per day22The “Biologics Boondoggle”“A breast cancer patient’s annual costfor Herceptin is $37,000…1Small moleculedrugsBiopharmaceuticalsPeople with rheumatoid arthritis orCrohn’s disease spend $50,000 a yearon Humira……and those who take Cerezyme to treatGaucher disease….spend a staggering$200,000 a year…“…the top six biologics already consume43% of the drug budget for MedicarePart B”1Source: NY Times, March 2010All trademarks are the property of their respective owners.6 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

© Sandoz 2012By 2016, 7 of the top 10 pharmaceuticalsworldwide will be biologics 1ProductType2016 Rev.(USD bn)1. HUMIRA Biologic 11.2 8.32. ENBREL Biologic 7.5 7.93. RITUXAN Biologic 7.4 7.14. AVASTIN Biologic 7.2 6.05. REMICADE Biologic 7.0 7.26. SERETIDE/ADVAIR Respiratory / device 6.8 8.17. HERCEPTIN Biologic 6.3 5.98. REVLIMID Small molecule 5.9 3.29. CRESTOR Small molecule 5.8 7.110. LANTUS Biologic 5.5 5.52011 Rev.(USD bn)1Source: Evaluate Pharma March 20 2012, vaccines excludedNote: All trademarks are the property of their respective owners.7 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

OverviewWhy biosimilars?QbD and the scientific approach to biosimilar developmentClinical trial requirementsSuccesful commercialization broadens patient access8 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

Biologics are more complex than small moleculesand mAbs more complex than simple biologicsAspirin®small chemical moleculeMolecular weight= 180 Daltons0 amino acidsCalcitoninsimple biologicMolecular weight= 3,455 Daltons~ 32 amino acids- w/o host cell modifications- produced in yeast, bacteriaMonoclonalAntibody (IgG)complex biologicMolecular weight= 150,000 Daltons~ 1300 amino acids- w/host cell modifications(glycosolations, etc)- produced in mammalian cells9 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

Innovation required in both technical developmentand clinical developmentKey challengesTime &Investment• Significant expense (USD 100 - 250m)• Long time to develop (7-8 years)TechnicalDevelopment• Achieving “highly similar” to match originatormolecule profile• Matching final dosage form of originatorClinicalDevelopment• Use of novel endpoints and populations to confirmbiosimilarity (not de novo safety/efficacy)• Clinical trial design to support extrapolation acrossindications, interchangeability & commercial success10 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

Biosimilars must be systematically engineered tomatch the reference productReferenceproductClinical2. Confirmationof biosimilarity1. Target directeddevelopmentTarget rangeDrug productdevelopmentPK/PDPreclinicalPurification processdevelopmentProcessdevelopmentBiologicalcharacterizationBioprocess developmentRecombinant cell line developmentAnalyticsPhysicochemicalcharacterizationLeveraging biological variability11 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

“Acceptable changes in quality attributes ofglycosylated biopharmaceuticals”2,01,61,20,80,40,0Unfucosylated G0[% of glycans]08.2007 12.2008 05.2010 09.2011Expiry Date1401201008060Pre-ShiftADCC Potency[% of reference]Pre-ShiftPost-ShiftPost-Shift08.2007 12.2008 05.2010 09.2011Expiry DateSchiestl, M. et al., NatureBiotechnology 29, 310-312, 2011)• Monitoring batches of anapproved mAb revealed ashift in quality• Shift in glycosylation(structure) pattern resultsin different potency incell-based assays(function)• Indication of a change inthe manufacturingprocess• Sandoz observed suchshifts in several originalproductsDifference to post-change versionsometimes greater than to biosimilar12 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

QbD biosimilar product specifications impacted byvariability in originator product• Analytical methods are sensitive to differentiate betweenBatch to batchBatches before and after a change of the manufacturing processBatches from different sites• Analytical methods candetermine whether batchessourced in different countriesare identical or notMicroheterogeneity of protein structure» Purity profiles» Glycan distribution% G2F605040302010Enbrel ® batches - G2F amountsUS LyoUS LiqEU US Lyo LiqEU LyoEU Liq LyoEU LiqEU LiqManufacturingprocess changeSchiestl, M. et al., NatureBiotechnology 29, 310-312, 2011)001.2007 02.2008 03.2009 05.2010 06.2011Expiry dateAll trademarks are the property of their respective owners.13 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

ADCC (%of Reference)Quality Attribute [%]Structure function relationships refined in biosimilardevelpopment: Adjusting ADCC in clone selection7006005004001086Screening of bioreactorconditions300200100420Range of orginatoron market toonarrow to deduceS/F-relationship0 2 4 6 8bG0-F [rel. %]ParentalCell LinePoolsVariability observed during cellline development enableselucidation of quantitative S/Frelationship0Clones Clones Clones Final CloneCell Line X Cell Line X Cell Line X Cell Line XPool A Pool A Pool B Pool BFrom: McCamish, Clinical Pharmacology & Therapeutics, Feb 201214 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

Fingerprint mAbs/Fusion proteins as FDA might see itPrimary structure e.g.:LC-MS intact massLC-MS subunitsPeptide mappingImpurities e.g.:CEX, cIEF acidic and basic variantsLC glycationPeptide mapping deamidation,oxidation, mutation, glycationSEC/FFF/AUC aggregationBiological activity e.g.:Binding assayADCC assayCDC assay15 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012Higher order structure e.g.:NMRCD spectroscopyFT-IRPTMs e.g.:NP-HPLC-(MS) N-glycansAEX N-glycansMALDI-TOF N-glycansHPAEC-PAD N-glycansMALDI-TOF O-glycansHPAEC-PAD sialic acidsRP-HPLC sialic acidsCombination of attributes e.g.:MVDA, mathematical algorithmsA comprehensive set and combination of orthogonal analytical methodsrevealing structure-function relationships, delivering in depth comparabilityinformation and allowing extrapolation towards non-measured attributes

Residual uncertainty decreasesOverview of FDA Approach to BiosimilarityTotality of evidence, stepwise, and risk based approachClinicalScope and magnitude depends on extent ofresidual uncertainty from below steps- No need to independently establish safety or efficacy- Immunogenicity data is minimally expectedPK/PDPK and PD (where there is a relevant PD measure)studies are generally expectedPreclinicalFlexibility regarding need for animal studies- Animal toxicity studies may not be warranted- Useful if safety uncertainties remain before first-inmanstudiesBiologicalcharacterizationPhysicochemicalcharacterizationAnalytical characterization is the foundation- The more comprehensive and robust the data, thestronger the justification for selective and targetedapproach to animal and human testingUnderstanding of reference product is important:MOA, SAR, clinical knowledge, availability of clinicallyrelevant PD measure, etc.16 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

OverviewWhy biosimilars?QbD and the scientific approach to biosimilar developmentClinical trial requirementsSuccesful commercialization broadens patient access17 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

FDA: Abbreviated clinical development feasibleSource: Steven Kozlowski, Director, Office of Biotechnology Products, Biosimilar Workshop, APEC, Seoul, April 4, 201218 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

Exaggerated expectations for Biosimilar clinical trials• Temple: “Recent biosimilar legislation calling for biosimilar products to beshown to have ‘no clinically meaningful differences’ for the reference productwill greatly extend discussion of how to determine when two treatments are‘essentially’ the same. At present, however, there is no real consensus orstandard.”• “With rare exceptions, differences between drugs, if any, will be small,considerably smaller than the whole effects of the drugs, which themselvesare often small. And the difference one would want to rule out is also small.”• If it is challenging to differentiate between two distinct therapeutic entities, itwill be even more difficult to differentiate between a biosimilar and originatoreven with minor differencesRobert Temple: Clinical Trials 201119 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

ACR20 Response Rate [%]Clinical trials are not even sensitive enough to differentiatedifferent biologics: Anti-TNFs‣ Response Rates of anti-TNFs vary depending on studyprotocols20 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

SPR real time binding assay much more sensitive todiferentiates anti-TNFs and demonstrate biosimilarityKaymakcalen, et al: Clinical Immunology, (2009) 131, 308-316www.diahome.org21 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

Sandoz biosimilars created to match originator bindingkinetics AND clinical performanceSandoz internal data:OM = originator materialPIL = pilot scaleConf = clinical scalemAB3 = different anti-TNF mAb22 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, September 2012

OverviewWhy biosimilars?QbD and the scientific approach to biosimilar developmentClinical trial requirementsSuccessful commercialization broadens patient access23 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

Sandoz Biopharmaceuticals: Our Vision“We will dramaticallyincrease worldwidepatient access toessential, high qualitybiopharmaceuticals byadvancing our globalleadership position inbiosimilars”24 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

UK example:Biosimilars expand access to G-CSF 1UK G-CSF volume growthPercent change vs. previous yearSep. 2008Biosimilarsapproved2007200820092010Sandoz’ filgrastim is not approved in the US.2011• G-CSF prevents hospitalre-admissions due to infections• Many physicians have movedG-CSF back to 1 st -line cancertreatment due to lower biosimilarscost• Sandoz’s filgrastim (G-CSF)“Patient Support Kits” expandpatient access:– Patients self-administer at home– Substantial efficiency savings1Granulocyte colony stimulating factorSOURCE: IMS, NHS25 | Genetic Engineering & Biotechnology News Webinar on Biosimilars, November 2012

Learning from History: Making Biosimilars aRealityGraham Jones, Ph.D.Chair, Department of Chemistry & Chemical BiologyNortheastern University

Analytical Characterization of Biopharmaceuticals andDetermination of BiosimilarityProfessor Graham B. JonesNortheastern UniversityGEN Webinar11/1/12

ioprocessreactors – allshapes and sizesc.f. small molecule manufacturing- contaminants- viruses- excipients/nutrients- formulants in solution, half life- genetic ‘drift’ possible

Amino acids ->peptides->proteins->PTM’sGlycosylationGalactosylationFucosylationhigh mannose derivativessialylationprocessingoxidation,phosphorylation,sulphation,lipidation,disulphide bond formationdeamidationposttranslationalmodifications

alpha helicesProtein Structurestertiary structurebeta sheetsquaternary structure

Tissue PlasminogenActivator [tPA]: anti bloodclotting agent [e.g.alteplase, reteplase,tenecteplase] 527 aminoacid protein with 17 S-Sbridges and 3glycosylation sitesOxidation, deamidation,glycosylation, proteolysis,sequence variation, chainisoforms, andcarbohydrate derivatives= 1.09 x 10 9 possiblevariants!!

Mab’s pose even greater challenges: Variation of a single fucose (mass=164)can have an immediate impact on activity of IgG (mass ~150,000)!www-immuno.path.cam.ac.uk/~mrc7/

Role of appropriate due diligence studies invalidated methods development: the Heparin crisis….• 81 deaths in US, 100’s worldwide• Contaminant – semi synthetic oversulfatedchondroitin sulfate• Analysis by NMR, CE, LC/MS (enzymaticcleavage)• Culprit determined by NMR

Role of appropriate due diligence studies in validated methods developmentEprex- pure red cell aplasia: analytical standards for analysis of leachates [HSA substitutepolysorbate 80]-Retacrit and Silapo: O-glycan chains and variant of sialic acid detected - different [canine]immunogenicity profiles- Abseamed, Binocrit and Epoetin alfa Hexal: IAC shows different glycosylation patterns viz.higher levels of high-mannose-type structuresKey: EPO is not a single active ingredient – is a heterogenous mix of up to 14 differentisoforms! Isoelectric Focusing analysis of two isoforms shows similar patterns and sialicacid content but very different biological activity e.g in vivo activity of 226,000 U/mg versus400,000 U/mgBurg et al 1998 PCT/EP/98/07876

Underscoring the need for appropriate, harmonized analytics..cation exchange chromatography reveals additional information

An Open Forum on the Scientific and Regulatory Issuesof Biosimilars and Follow-on BiologicsBoston, MA USACopley Marriott Hotelwww.barnett.neu.edu/biogenerics/Address by:Senator Edward KennedyChairman of the Senate Health,Education, Labor and PensionsCommitteePlenary by:Dr. Randall LutterActing Deputy Commissionerfor Policy, US FDAA high level meeting of the international biotechnology and generics industries, togetherwith government regulators and academic laboratories, for an open forum and collegialdebate on the scientific and regulatory issues in the introduction of generic complexbiological drugs.For additional information or to receive updates, email Biogenerics2008@neu.eduOrganizing Chair:Prof. Barry KargerDirector, Barnett Institute, andJames L. Waters Chair,Northeastern UniversityHosted by:

Biologics Price Competition and Innovation Act (BPCI Act), enacted March 2010The FDA sought comments on the following issues related to determiningbiosimilarity:1. What scientific and technical factors should the agency consider indetermining whether the biological product is highly similar to the referenceproduct notwithstanding minor differences in clinically inactive components?2. What scientific and technical factors should the agency consider indetermining the appropriate analytical, animal, and clinical study or studies toassess the nature and impact of actual or potential structural differencesbetween the proposed biosimilar product and the reference product?3. What range of structural differences between a proposed biosimilarproduct and the reference product is consistent with the standard “highlysimilar” and may be acceptable in a 351(k) application if the applicant candemonstrate the absence of any clinically meaningful differences between theproposed biosimilar product and the reference product?4. Under what circumstances should the agency consider finding that animalstudies or a clinical study or studies are “unnecessary” for submission of a351(k) application?

ICH Harmonized Tripartite Guideline “Q5E” – [ICH working group, Nov. 2004]“Comparability of Biotechnological / Biological Products Subject to Changes in theirManufacturing Process”Excerpts:1. Physicochemical Properties [ICH Q6B][molecular homo/heterogeneity, higher order structural analysis –secondary, tertiary,quaternary structures]2. Biological Activity3. Immunochemical Properties4. Purity, Impurities and Contaminants[isoforms, degradation products]Analytical TechnologyPost translational modificationsStructural differencesIsomerizationO-glycan characterizationHigher order structuresMonomeric and dimeric forms

Nature Reviews Drug Discovery, 2012, 11, 527-540

FDA desire for specific innovations in biotherapeuticanalysisSteven Kozlowski, M.D.Director, Office of Biotechnology ProductsThree Specific Properties Needing Improved MeasurementFDA has identified three properties of therapeutic proteins that cannot besufficiently measured at this time but that are very important for understandingthe behavior of protein drugs. Improved analytical methods to measure these threeproperties would be particularly useful in determining the extent of similarity ofbiological protein products intended to be similar.1. Post-translation Modifications …2. Three-dimensional Structure …3. Protein Aggregation …TESTIMONY BEFORE THESUBCOMMITTEE ON TECHNOLOGY AND INNOVATIONCOMMITTEE ON SCIENCE AND TECHNOLOGYU.S. HOUSE OF REPRESENTATIVESSEPTEMBER 24, 2009

FDA desire for specific innovations in biotherapeuticanalysisSteven Kozlowski, M.D.Director, Office of Biotechnology Products1. Post-translation ModificationsAs indicated previously, proteins contain added structural features, such asattached sugar chains, that may be critical for their clinical activity. Theseattached modifications can be complex and heterogeneous, and wecurrently lack standardized analytical methods to qualitatively andquantitatively assess the structure as it relates to the intact protein andunderstand the relationship of the modifications to potency and clinicalperformance. We are particularly interested in better methods foranalyzing the sugars (glycosylation) and other modificationsknown to affect the medicinal activity of these products.TESTIMONY BEFORE THESUBCOMMITTEE ON TECHNOLOGY AND INNOVATIONCOMMITTEE ON SCIENCE AND TECHNOLOGYU.S. HOUSE OF REPRESENTATIVESSEPTEMBER 24, 2009

HPLC-Chip Platform for Nanospray LC/MS:Agilent TechnologiesHPLC-Chip/MSinterface1200 NanoLC System6000 Series Mass Spectrometer (Iontrap, SQ, QQQ, TOF, Q-TOF)

Differences in Sites of Fucosylation Can be Measured byMS-MS [CID, ECD, ETD]B)x10 657.2424rFIX4: TOF MSMS 1901.60ES+2.15e3Tag454.1565(M+2H)2+366.1502204.0931488.21902 .7115x10 657.2350pd-FIX7: TOF MSMS 1901.60ES+294366.1260204.0841 803.26651688 .5410200 400 600 800 1000 1200 1400 1600 1800m/zNature Reviews Drug Discovery, 2012, 11, 527-540

Relative AbundanceLC-MS-NMR and micro-NMR:-analysis of heparin O-glycans1008015.80458.5815.95458.67base peak60402001003.91433.585.64433.5815.05427.5813.60574.0817.02711.5817.38972.4217.77527.0023.85725.2525.01580.1728.04792.8329.43843.588060Hex 5 GlcNac 240micro–NMR has high20sensitivity for LC-isolatedanalytes 06.72972.0810.39972.1713.66972.0816.32972.7517.2918.04972.5821.84972.5024.81972.3327.41972.9230.62972.58Tryptic GlycopeptideStructural Reporter Groups Resolved(RNase B shown)NMR solved the Heparin Crisis 11 Guerrini … Sasisekharan. Nat. Biotech 26(6) 669-675 (2008)

ETD to analyze formation of isoAsp [isomerization of Asp /deamidation of Asn]: amyloid bisoAsp can beimmunogenic0.1Enrichment of isoD by Asp-N digestionbefore digestionafter digestion0.08Maximum, 100% isoDz 10-57/z 100.060.040.02Anal.Chem., 82, 7485 (2010).00% 1.9% 3.7%Spiked isoD percentage

FDA desire for specific innovations in biotherapeuticanalysisSteven Kozlowski, M.D.Director, Office of Biotechnology Products2. Three-dimensional StructureAs described previously, proteins must be folded into a three-dimensionalstructure to become functional (sometimes a three-dimensional structure can bemisfolded). The proteins within a biologic will have one major three-dimensionalstructure along with a distribution of other variants differing in three-dimensionalstructure. Our current ability to predict the potency of biologicswould be enhanced if we had improved ability to measure andquantify the correct (major) three-dimensional structure, aberrantthree-dimensional structures (misfolding), and the distribution ofdifferent three-dimensional structures.TESTIMONY BEFORE THESUBCOMMITTEE ON TECHNOLOGY AND INNOVATIONCOMMITTEE ON SCIENCE AND TECHNOLOGYU.S. HOUSE OF REPRESENTATIVESSEPTEMBER 24, 2009

High order structures ofvery complex proteins are now possible….Complete disulfidemapping – 17disulfidesETD/CID MSPreviousassignments byhomology to otherproteins…Anal. Chem., 82,5296 (2010)

Three dimensional structural interrogation using HDX-MSAmide hydrogen exchange in backboneD2OTime-course analysis permits assessmentof structure and dynamics

Ability to assign structural changesDecrease indeuteriumincorporation indeglycosylatedformIncrease indeuteriumincorporation indeglycosylatedformRegion lessprotected when IgGis deglycosylatedNature Reviews Drug Discovery, 2012, 11, 527-540Region moreprotected whenIgG isdeglycosylated

FDA desire for specific innovations in biotherapeuticanalysisSteven Kozlowski, M.D.Director, Office of Biotechnology Products3. Protein AggregationSome biological products can stick to one another. When many proteinmolecules stick together, they are referred to as aggregates and have thepotential to cause adverse immune responses in patients. There are many formsand sizes of aggregates and many current methodologies have gaps in theirability to detect different types of aggregates. Our ability to minimizeadverse immune reactions would be enhanced if we had improvedability to measure and quantify different types of aggregates.TESTIMONY BEFORE THESUBCOMMITTEE ON TECHNOLOGY AND INNOVATIONCOMMITTEE ON SCIENCE AND TECHNOLOGYU.S. HOUSE OF REPRESENTATIVESSEPTEMBER 24, 2009

Nature Reviews Drug Discovery, 2012, 11, 527-540IMS separation of monomeric insulin fromdimeric insulin6+Intensity3+637 Å 26+1231 Å 23+Drift Time (ms)

Analysis of human insulin analogs by IMS-MSHumulinHumalogApidraLantus Novolog Levemir

size exclusionchromatography (SEC)analyticalultracentrifugation (AUC)Other methods for analysisof aggregates and SVP’s [AF4,AUC, LS do not alter sample]asymmetric flow fieldflow fractionation (AF4)dynamic lightscattering (LS)

ReferencesAnalytical tools for characterizing biopharmaceuticals andthe implications for biosimilars. Berkowitz, Steven A.;Engen, John R.; Mazzeo, Jeffrey R.; Jones, GrahamB. Nature Reviews Drug Discovery, 2012, 11, 527-540.The Forward Path for Biopharmaceuticals and Biosimilars:Emerging Options in the Selection of Host Cell Systems.Wager, Krista, Jones, Graham, Current Biotechnology,2012, 1 (4), 297-317.

Learning from History: Making Biosimilars aRealityMarie Rock, Ph.D.Senior Scientific Director of Large Molecule BioanalysisWIL Research

Regulatory Expectationsfor BiosimilarsMarie Rock, Ph.D.

Are the Molecules Similaror are They Different?Is it what I am seeing…ORIs it what I am seeing with?

http://bit.ly/Biosimilars_Guidance

Focus of GuidanceQualityScientificSource: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product

Approaches to Demonstrating BiosimilarityFDA considers the totality of theevidence provided to support ademonstration of biosimilarityFDA recommends the use of astepwise approach in thedevelopment of biosimilar productsSource: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product

A Stepwise Approach to Demonstrating BiosimilarityGeneralscientificprinciples inconductingcomparativestructural andfunctionalanalysisAnimalTestingHumanPK andPDStudiesClinicalSafetyandEfficacyClinicalImmunogenicityAssessment

Clinical Studies are Required to DemonstrateBiosimilarity“…current analytical methodology may not be able todetect all relevant structural and functional differencesbetween two proteins. Thus…data derived from analyticalstudies, animal studies and a clinical study or studies arerequired to demonstrate biosimilarity unless FDAdetermines an element is unnecessary.”**Section 7002(a)(2) of the Affordable Care Act, adding section 351(k)(2)(A)(i)(I) of the PHS ActSource: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product

Our Focus: PK & Immunogenicity•Our focus: using immunochemical techniques to showbiosimilarity in vivo.•If the comparative PK and immunogenicity betweentwo proteins are “similar” (clinically and non-clinically),then they are considered “biosimilar”.

COMPARATIVE PHARMACOKINETICS

Typical ELISA Method for PK DeterminationTMB SubstrateStreptavidin-HRPBiotinylated-Anti-Drug AntibodyBiologic in SampleCapture: Anti-DrugAntibody Reagent

Pharmacokinetics: Recommended Approach•ONE ASSAY:• Same Technology• Same Reagents• Same Conditions•One Calibration Curve•One set of controls for Originator•One set of controls for BiosimilarThe controlswill mimic thesamples

Best Practice for Side by SideComparisons for PKEvaluation of ResultsCompareDataPrecisionControls:AnalyticalRecoverySelectivityStudy Samples:Assessment of PKParameters

IMMUNOGENICITY

Progression of Immunogenicity Testing1956 2002 2004 2008 20092012Berson And YalowReport Insulin BindingImmunoglobulin in InsulinTreated PatientsNicole Casadevall, et. al.Report Anti-rHuEPO Absresulting inPure Red cell aplasiaAnd anti-EPO AbWhite PaperEMEARE: AntibodyScreening AssaysFDADRAFTFDADRAFT

DrugTherapeutics Can Induce Antibody Formation1000.0Antibody Negative Pt.5100.0410.01.0Antibody Positive Pt.32Antibody0.110.010 3 6 9 12 15 18 21 24 27 300

Antibodies Can Cross React with Multiple Antigens]FAB]Fc

Assay Formats: Antibody Detection/ScreeningAntibody DetectionMSD Bridging FormatECL ReadoutSulfo-Tagged -DrugAntibody in SampleAntibody DetectionMSD Direct FormatECL ReadoutSulfo-Tagged – AntiSpecies AntibodyBiotin-Tagged DrugAnti-Drug AntibodyStreptavidin CoatedPlatesDrug Coated Plates

Why Conduct an Immunogenicity Assessment?PurposeIncidenceSeverityImpactPKAnaphylaxisCross react withendogenouscounterpartSource: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product

Immunogenicity Testing Cascade:Compare Originator and BiosimilarAntibody screening assayAb PositiveCharacterize and TiterTest for NeutralizingAntibody Activity

Antibody Characterization: Binding andNeutralization AntibodiesSpecificityIsotypeTiterProgressionDurationClearanceClinical SafetySpecificityIsotypeTiterProgressionDurationClearanceClinical SequaleaEndogenousCounterparts

Best Practice for Side by Side Comparisons forAssessing ImmunogenicityScreening AssayPanelOne AnalyticalMethod• Detect antibodyresponse for bothwith same assay• Validate one assay

Cell Based Assays:Product Release and Neutralizing AntibodyDetectionAssessment of “Activity” (usebiosimilar release assay)Typically Product ReleaseAssay Adapted to perform inHuman Serum MatrixCan take 6 – 9 months todevelop and validateVery sensitive to changes inreagents and assayconditions

Cell Based Assay for Drug Bioactivity

Target Cell and Neutralizing Antibody

Summary•Comparative pharmacokinetic andimmunogenicity data will be critical inestablishing biosimilarity• Sound immunochemical analytical methods willbe the key to success•Requires in-depth understanding of the techniques•Careful attention to detail•Communicate early and often

Corporate OverviewIllinoisWIL Research Company is a leadinginternational provider of nonclinical development,safety assessment and regulatory consultingservices to the pharmaceutical, andbiotechnology industries.OhioThe NetherlandsClients count on WIL Research Company and its1000+ employees to:• Apply advanced scientific and technological expertise toassure the safety of new small molecule and biotherapeuticcandidates• Increase their development and compliance speed,precision and productivity5Antibody Titration Curves432101 10 100 1000 10000 100000 100000081

WIL: Analytical Laboratory with Experience:• Method development• Validation• Sample analysis• All analytical platforms• Cell based assays (potency and Nab)• QA Unit Current in Evolving Regulatory Requirements

Thank YouMarie Rock, Ph.D.

Learning from History: Making Biosimilars aRealityLearning from History: Making Biosimilars a RealityQ&A

Learning from History: Making Biosimilars aRealityYour ModeratorJohn SterlingEditor-in-ChiefGenetic Engineering & Biotechnology News

Learning from History: Making Biosimilars aRealityMark McCamish, M.D., Ph.D.Global Head of Biopharmaceutical DevelopmentSandoz Biopharmaceuticals

Learning from History: Making Biosimilars aRealityGraham Jones, Ph.D.Chair, Department of Chemistry & Chemical BiologyNortheastern University

Learning from History: Making Biosimilars aRealityMarie Rock, Ph.D.Senior Scientific Director of Large Molecule BioanalysisWIL Research

Learning from History: Making Biosimilars aRealityThank You For AttendingLearning from History: Making Biosimilars a RealityBroadcast Date: Thursday, November 1, 2012Time: 1:00 pm ET, 10:00 am PTSponsored by