Biosimilars- case studies (Analytical point of view)

BySada siva , Scientist,Analytical Development, Biologics/Bio-pharma professional.Sadasivaibt@gmail.comRole of Analytics in Biosimilars Approval(mAb Case sudy)

This Discussion covers.Quick introduction on Biosimilars and Regulatory approval bodies in India , Europe and USA

Regulatory guidelines, Analytical requirements and approaches used to show Biosimilarity

Biosimilar mAb case study: Approved in USA & Europe- Analytical data generated to show Biosimilarity- Physico chemical / structural analysis (not functional assay)

What are these Biosimilars

A 'biosimilar is a biological medicine that is similar to existing biological medicine authorised .Biosimilars/ Follow on biologics/similar biologics/subsequent entry biologics/Similar biotherupetic productFDA: Biosimilarity is defined in section 351(i) of the PHS Act to mean that the biological product is highly similar to the reference product notwithstanding minor differences in clinically inactive components and that 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

Why Biosimilars needed..??Bringing a new drug s into market s a very complex process which includes large no. of .non-clinical and clinical studies. It is associated with high developmental Cost and long time, Low success rates.Making Drugs cost higher to and low access to large patients.Regulators allow this to promote innovation in unmet medical needs.

To a word on why biosimilars concept is imp4

Evolution of the Concept of Abbreviated medicines / Generics/ BiosimilarityHence to overcome those constrains, Regulatorys came up wih Abbreviated pathway for copy /similar drugs with reduced design.( once the patent expire for new drugs)Generic small molecule drugs introduced sameness Biosimilarity (EU 2004, WHO 2009, US 2010) based on highly similar to the reference and no clinically meaningful differences

Differences in Data requirements between New and biosimilar molecules

Advantages of Biosimilars

Similar drugs having same efficacy and safety comes to market .Development cost and time are relatively less for BiosimilarsIt Helps in Cost reduction of DrugsIt also Increased access to Patients for costly treatments.

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Biosimilars approval in IndiaBiosimilars approval is regulated by Drug controller general India (DCGI) , Central Drugs Standard Control Organization (CDSCO) and the Department of Biotechnology (DBT)Guidelines on Similar Biologic: Regulatory Requirements for Marketing Authorization in India (2015)First approved Biosimilar: Biosimilars approved : GCSF, Rituximab, Interferons, Bevacizumab, erytrhopointion, Darbipoitin, Adalimumab, Etarnacept

Biosimilars approval in EuropeBiosimilars approval is regulated by european medicines agency (ema) Legal Basis: Directive 2001/83/EC First approved Biosimilar: Omnitrope (somatropin) from Eli Lilly (in 2006 )Others Biosimilars: Around 22 versonsGrowth hormone, EPO, GCSF, Infliximab, Eternacept, Insulin, Interferon Etc..

Biosimilars Data requirement in Europe

Biosimilars approval in Europe

Biosimilars approval in USABiosimilars approval is regulated by US FDA under PHS act.First approved Biosimilar: rGCSF from Sandoz (in 2015 )Others Biosimilars: Infliximab from Cellitron-First Biosimilar mAb in 2016Etarncept From Sandoz in 2016

Biosimilars approval in USA

Advantage of 351(K) Biosimilar PathwayQuality Considerations in Demonstrating Biosimilarity of a Therapeutic Protein Product to a Reference ProductScientific Considerations in Demonstrating Biosimilarity to a Reference Product

Infliximab (by cellitron)Case study

First Biosimilar mAb in US and Europe

Reference product: Remicade (Janssen )

One of the Top 10 revenue generating biologics.

Indications: Rheumatoid, Arthritis (RA), Psoriatic Arthritis (PsA), Crohns Disease (CD) etc.

Infliximab biosimilar- Regulatory History and FDA Interactions.Proposed Product development began in 2008 for Europe market, was guided by EMA biosimilar guidelines and scientific advice interactions.Based on these data, Proposed Product became the first licensed biosimilar monoclonal antibody in the world. Proposed Product is currently licensed for use in 67 countries.Following 2012 FDA draft biosimilar guidance, FDA provided input on the development program for Proposed Product at Biosimilar Biological Product Development meetings held on 2013 (BPD Type 3) and 2014 (BPD Type 4)

Infliximab biosimilar- Regulatory History and FDA Interactions.FDA suggested that adequate analytical and PK bridging data could be sufficient to demonstrate Biosimilarity between Proposed Product ,US Remicade and EU RemicadeCELLTRION conducted analysis of the structural and physicochemical attributes and testing of biological activities using US Remicade, EU Remicade and Proposed Product to demonstrate analytic biosimilarity In addition, CELLTRION conducted a 3-way PK Study to bridge the EU Remicade PK data with that of US Remicade.

Biosimilarity Approach. Biosimilarity is comparison exercise between a proposed and reference product.The aim of the Biosimilar exercise is to demonstrate that the Proposed product and the reference medicinal product are similar at the level of the finished product. It is not expected that all quality attributes of the biosimilar product will be identical to RMP. However, where qualitative and/or quantitative differences are detected, such differences should be justified and, where relevant, demonstrated to have no impact on the clinical performance of the product.

Analytical comparison plays main basis for Biosimilarity.Results of analytical comparison , helps to decide further to the number of of Non clinical and clinical comparability studies required to prove Biosimilarity.

Factors for consideration in assessing Biosimilarity (FDA).Reference Product and Reference StandardsExpression systemManufacting processAssessment of Physicochemical PropertiesFunctional ActivitiesReceptor Binding and Immunochemical PropertiesImpuritiesStability

Reference medicinal Product (RMP).RMP used in Biosimilarity should be approved and licensed by respective regulatory authority.Single sourced RMP should be used all the studies (Quality, Non Clinical and Clinical studies )However, with the aim of facilitating the global development of Biosimilars , it is allowed to compare the biosimilar with a other regulatory authorised comparator. In this Case , A bridging comparability study (Analytical and PK and or PD ) has to be performed between the two RMP products along with Proposed Biosimilar product.

Reference Medicinal Product.The reference medicinal product details should be captured properly (Complete details like lot, source, presentation)Multiple different batches of the reference medicinal product should be used and justified.The age of the different batches of reference medicinal product (Wide range over expiry is recommended )Publicly available reference standards (e.g. Ph. Eur.) cannot be used as the reference medicinal product for demonstration of biosimilarity

Reference Medicinal product Details

Side by side analysis performed between the 3 products in all the testsComparative stabilities performed.

SamplesNumber ofBatchesAge at the AnalysisProposed product7-239 -21 MUS Remicade7-322 29 MEU Remicade6-324-36 M

Expression system.

Careful selection should be done to produce High similar product .Expression system differences may results in undesired consequences like atypical Glycosylation pattern, a different impurity profile compared to RMPBetter to choose the same expression system

Manufacturing process.Develop a process to produce a Product highly similar to RMPQuality Target for product profile (QTPP) should be established at the early Development phase by RMP analysis and publicly available in formation.Performance and consistency of the manufacturing process of he biosimilar .Proper formulation should be selected .better to have the same as RMP does not need to be identical.If a different formulation and/or container & closure system , its impact on safety and efficacy should be justified

Assessment of Physicochemical Properties.Extensive State of the art analytical tests should be used to show Biosimilarity between the proposed product and the reference product.Lots used for the analyses should support the Biosimilarity of both the clinical material used in the clinical study(ies) intended to support a demonstration of BiosimilarityMultiple representative lots should be used understand the lot-to-lot variability of both productsA risk-based assessment should be performed to understand the impact and criticality of each quality attribute on safety and efficacy.

Extensive analytical testing should cover.Primary structures, such as amino acid sequence Higher order structures, including secondary, tertiary, and quaternary structure (including aggregation)Enzymatic posttranslational modifications, such as glycosylation and phosphorylation Other potential variations, such as protein deamidation and oxidation Intentional chemical modifications, such as PEGylation sites and characteristics

Analytical methods selectionSelection of tests chosen for comparability is mainly based on characteristics of the protein product and its potential impurities.Method to be used should have appropriate accuray, sensitivity and specificity to provide meaningful information as to whether the proposed product and the reference product are highly similar.Unlike routine quality control assays, tests used to characterize the product do not necessarily need to be validated. Qualification is needed for characterization tests.

Analytical methods selectionIt is often necessary to apply more than one analytical procedure to evaluate the same quality attribute. Because they provide independent data to support the quality of that attribute (e.g., orthogonal methods to assess aggregation). In addition, the use of complementary analytical techniques in series is encouraged , should provide a meaningful and sensitive informaion for comparing products. (such as peptide mapping combined with mass spectrometry of the separated molecules)

Product specific tests

Risk and criticality analysis.

Primary Structure analysis.Peptide mapping by HPLC showedt hat Proposed Product has a highly similar chromatographic peak profile to US and EU Remicade . No missing or additional significant peptides were detected and peaks had comparative retention times.

Structure analysis-Data.

Charge analysis.The same charge variant peaks were detected in Proposed Product, US Remicade, and EU Remicade. Proposed Product contains higher levels of Peak 1 and Peak 4 than US or EU Remicade. The difference is attributable to C-terminal lysine variability.Further analysis of the impact of C-terminal lysine variability was obtained from in vitro studies of biological activities, which showed that C-terminal lysine variability had no bearing on biologicalactivities in vitro. Additionally, incubation with IgG-free human serum resulted in rapid clipping of C-terminal lysine residues shown as reductions in Peaks 4, 5, and 6 which contain forms with one or two C-terminal lysine residues

IEX profile.

Glycan analysis.Glycan site analysis by LC-MSGlycan analysis by NP-HPLCSialic acids (Charged glycans) analysis by HPAEC-PADMonosaccharide analysislower amounts of afucosylated glycans (G0 and Man5) in Proposed Product had previously been observedAs there is no association of G0 with immunogenicity and G0 is present on endogenous antibodies, there appears to be no safety impact of the lower level.

Glycation.Although high similarity in percent glycation of Proposed Product and Remicade was notobserved as Proposed Product contained higher levels of glycated formsNo differences in biological activities between Proposed Product and Remicade are expected due to glycation, as was supported by data from similarity studies of biological assays including TNF binding and neutralization assays. Data from samples of Proposed Product and US Remicade with artificially created levels of glycation showed that glycation has no impact on FcRIIIa binding affinity of Proposed Product or US Remicade.No impact on immunogenicity is expected as antibodies are glycated on incubation with serum and in vivo

Impurities..The sponsor should characterize, identify, and quantify impurities in the proposed product and the reference product, to the extent feasibleIf a comparative physicochemical analysis reveals comparable product-related impurities at similar levels between the two products, no additional studies required.If proposed product different or higher levels of impurities than in the RMP, then additional pharmacological /toxicological or other studies may be necessary to understand their impact.

Impurities..It is preferable to rely on purification processes to remove impurities . . . rather than to doing additional tesing.Differences that may have a safety advantage (e.g. lower levels of impurities) should be explained.The process-related impurities in the proposed product are not expected to match those observed in the reference product. But they should be minimised by process.The potential impact of the differences in the impurity profile upon safety should be addressed and supported by appropriate data.

Aggregate analysisSEC-HPLC detects monomer, molecular weight forms(HMW). Proposed Product lots had a higher HMW content (mean value 0.8%) than US &EU Remicade lots (0.2%)The clinical relevance of the higher HMW and lower monomer content in relation to immunogenicity was assessed through the repeat-dose studies in RA and AS and was shown not to be clinically meaningful.SEC-MALS also detects monomer, HMW, and their sizes.Analytical Ultracentrifugation (AUC) is an alternative method to measure monomer and HMW forms. All 3 products contained monomer as the predominant sedimenting species.

Stability Data.Proposed Biosimilar & RMP degradation profiles should be established .Accelerated and stress stability studies, as well as forced degradation studies should be performed side by side.These comparative studies should be conducted under multiple stress conditions (e.g., high temperature, freeze thaw, light exposure, and agitation)Results should be presented and if these studies may reveal product differences that warrant additional studies .Shelflife should be determined from the Realtime stability data of the Proposed Biosimilar and extrapolation is no recommended. (RMP in real time stability is not required for Biosimilarity)

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Stability studies performed

Functional ActivitiesFunctional assays are imp part of Biosimilarity as an essence step establishing complete characterization profile. These tests act to complement physicochemical analyses and are a qualitative measure of the function of the protein product. For complex proteins, the available analytical technology, the physicochemical analysis may be unable to confirm the integrity of the higher order structures. In his case functional assays are very helpful.

Receptor Binding PropertiesReceptor Binding and Immunochemical PropertiesWhen binding or immunochemical properties are part of the activity attributed to the protein product, analytical tests should be performed to characterize the proposed product in terms of these specific properties, (e.g., if binding to a receptor is inherent to protein function, this property should be measured and used in comparative studies)

Statistics recommended

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Some important points to consider about Biosimilars are. Biosimilars dont have to be proved for every indication.Other than looking at the guidelines there are no requirements for development / trials of biosimilars.Biosimilars must have comparable quality, safety and efficacy.Each Biosimilar will have gone through the appropriate regulatory pathway.Every Biosimilar product is different and each should be considered an individual brand in their own right.Because every biosimilar is different no one set of regulatory rules will work for all developing the need for each to be considered on a case by case basis.

Other related topics for discussion.

Functional studies comparison of this case studyNonclinical and Clinical studies of this case studyOther Biosimilar Case studies (GCSF and Etarnacept)

Learning starts from questioning ??????????

For giving this opportunity

References.EMEA:Guideline on similar biological medicinal productsEMEA:Guideline on similar biological medicinal products containing biotechnology-derived proteins as active substance: non-clinical and clinical issuesEMEA:Guideline on similar biological medicinal products containing monoclonal antibodies non-clinical and clinical issues) FDA:Quality Considerations in Demonstrating Biosimilarity of a Therapeutic Protein Product to a Reference ProductFDA:Scientific Considerations in Demonstrating Biosimilarity to a Reference Product FDA:Clinical Pharmacology Data to Support a Demonstration of Biosimilarity to a Reference ProductICH guideline Q5E: Note for guidance on biotechnological/biological products subjected to changes in their manufacturing processICH guideline Q5C: Note for guidance on quality of biotechnological products: Stability testing of biotechnological/biological productsICH guideline Q6B: Note For Guidance on Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological ProductsIndia DCGI: Guidelines on Similar biologics by CDSCO and DBT