Fresenius Kabi has developed FKS518, a fully human monoclonal antibody biosimilar to denosumab. The clinical development program included two randomized comparative trials: a PK study in healthy volunteers and a safety and efficacy study in osteoporosis patients. The demonstration of similarity and equivalence between FKS518 and reference denosumab included the quantitation of the serum biomarker C-terminal cross-linking telopeptide of Type 1 collagen (CTx-1), a well-established marker of bone resorption. This paper details the development, validation, and analytical performance of the method for CTx-1 quantitation, emphasizing how the Context of Use (CoU) shaped the validation requirements. Application of the method in samples from the pharmacokinetic (PK) equivalence study allowed demonstration of pharmacodynamic (PD) similarity between FKS518 and Prolia. This publication also addresses the use of endogenous serum control (ESC) samples in monitoring assay performance. A retrospective analysis indicated that applying more flexible ranges and/or omitting ESC-based criteria for run acceptance would not have substantially changed the CTx-1 results. While recognizing the value of ESCs for stability and trending analysis and the importance of rigorous biomarker method development and validation in the assessment of biosimilarity, this paper fosters the discussion whether run acceptance based on tight ESC acceptance limits is always necessary.

BACKGROUND:Fresenius Kabi developed FKS518, a fully human monoclonal antibody biosimilar to denosumab, that inhibits osteoclast activation by targeting the receptor activator of nuclear factor kappa-Β ligand (RANKL). RANKL exists as a trimeric soluble protein (sRANKL) with high affinity for denosumab. Post-dose, sRANKL levels can rise due to drug-target complex accumulation, creating a risk of interference in bridging anti-drug antibodies (ADA) assays, particularly after acid dissociation. METHODS:We evaluated sRANKL interference in the ADA bridging assay and, to competitively block sRANKL, we introduced a specificity tier by adding osteoprotegerin (OPG). This approach enabled reanalysis of previously ADA-positive samples to confirm whether signals represented true ADA responses or artifacts caused by sRANKL interference. RESULTS:Acid dissociation significantly exacerbated target interference, resulting in ADA positivity rates of ~96-98% in clinical studies. Introducing a specificity tier corrected incidence to ≤3.9%. OPG incorporation did not change the minimum required dilution (MRD) of the assay and did not affect signals for negative/positive control, confirming assay integrity. DISCUSSION:These findings underscore the importance of early interference assessment and mitigation. A multi-tiered strategy, encompassing screening, confirmatory, and specificity tiers, provided a robust solution applicable to programs facing similar challenges.

Abstract Background/Aims Tocilizumab is an anti-interleukin-6 receptor monoclonal antibody indicated for treating rheumatoid arthritis (RA) and other inflammatory diseases. MSB11456 is a proposed biosimilar to US-licensed tocilizumab and EU-approved tocilizumab. It has already shown equivalent pharmacokinetic, pharmacodynamic safety, tolerability, and immunogenicity profiles to these products given subcutaneously (SC) as a single dose in healthy volunteers. This Phase III, multi-centre, randomised, double-blind, multiple fixed-dose, parallel group study compared efficacy, safety and immunogenicity of MSB11456 and EU-approved tocilizumab administered SC in moderate-to-severe RA patients. Methods Patients were randomised to 162mg MSB11456 or EU-approved tocilizumab for 24 weeks (W). At W24, patients receiving EU-approved tocilizumab were re-randomised to continue treatment or switch to MSB11456 for up to W52. Safety evaluations were conducted up to W63. A change from baseline in Disease Activity Score-28 Joint Count-ESR (DAS28-ESR) at W24 was analysed, as the primary efficacy endpoint, using analysis of covariance to determine the least squares mean (LSM) difference between MSB11456 and EU-approved tocilizumab; equivalence was considered if the 90% confidence interval (CI) was entirely within the FDA equivalence interval -0.6 to 0.5. Secondary endpoints were 20% improvement in ACR core set measures at W24 and DAS28-ESR at W12. Additional endpoints included ACR50/70, change in DAS28-CRP, Simplified and Clinical Disease Activity Indexes, evaluation of immunogenicity up to W55 and safety up to W63. Results Clinically relevant LSM decreases from baseline in DAS28-ESR were observed from W2 up to W24 with both treatments. The 90% CI for LSM difference in the change from baseline in DAS28-ESR between treatments was fully included within the predefined equivalence interval, which demonstrated therapeutic equivalence of MSB11456 and EU-approved tocilizumab (Table 1). Other efficacy endpoint analyses supported this conclusion. No discernible patterns in the nature or frequency of treatment-emergent adverse events (TEAEs) were identified to suggest a difference between drugs. Anti-drug antibodies incidence was similar among treatment arms. Switching from EU-approved tocilizumab to MSB11456 had no clinically relevant impact on efficacy or safety. Conclusion Equivalent efficacy, immunogenicity and safety profiles of MSB11456 and EU-approved tocilizumab were demonstrated in moderate-to-severe RA patients, confirming equivalence between MSB11456 and the EU-approved tocilizumab. Disclosure A. Zubrzycka-Sienkiewicz: None. M. Misterska-Skora: None. M. Socik-Pojawa: None. K. Klama: None. M. Ullman: Other; Employee of Fresenius Kabi SwissBioSim - Sponsor of the study. C. Petit-Frere: Other; Employee of Fresenius Kabi SwissBioSim - Sponsor of the study. A. Illes: Other; Employee of Fresenius Kabi SwissBioSim - Sponsor of the study. P. Baker: Other; Employee of Fresenius Kabi SwissBioSim - Sponsor of the study. J. Monnet: Other; Employee of Fresenius Kabi SwissBioSim - Sponsor of the study. J. Morais: Other; Employee of Fresenius Kabi SwissBioSim - Sponsor of the study. J. Brzezicki: None.

Objective To evaluate the efficacy, immunogenicity and safety of the proposed biosimilar MSB11456 versus European Union (EU)-approved tocilizumab reference product in patients with rheumatoid arthritis (RA) in a multicentre, randomised, double-blind, multinational, parallel-group study ( NCT04512001 ). Methods Adult patients with moderate-to-severe active RA and inadequate clinical response to ≥1 disease-modifying antirheumatic drug (synthetic or biologic) receiving methotrexate were randomised to receive 24 weekly subcutaneous 162 mg injections of either MSB11456 or EU-approved tocilizumab. Equivalence between treatments was considered if the 95% CI (European Medicines Agency)/90% CI (US Food and Drug Administration) for the difference in mean change from baseline to week 24 in Disease Activity Score-28 Joint Count with erythrocyte sedimentation rate (DAS28-ESR) between treatments was entirely within prespecified equivalence intervals (−0.6 to 0.6 and −0.6 to 0.5, respectively). At week 24, patients were rerandomised to continued treatment or MSB11456. Secondary efficacy endpoints to week 52, and safety and immunogenicity to week 55 were also evaluated. Results At week 24, the least squares mean difference in the change from baseline in DAS28-ESR between treatments was 0.01 (95% CI −0.19 to 0.22) in the 604 randomised patients. Similarity between treatments was shown for all other efficacy, safety and immunogenicity endpoints, including in patients who switched from EU-approved tocilizumab to MSB114466. Conclusions Therapeutic equivalence was demonstrated for efficacy endpoints, and safety and immunogenicity analyses support the similarity of the two treatments. The results of this study strengthen the evidence that the proposed biosimilar MSB11456 and EU-approved tocilizumab exert similar clinical effects.

The 17th Workshop on Recent Issues in Bioanalysis (17(th) WRIB) took place in Orlando, FL, USA on June 19-23, 2023. Over 1000 professionals representing pharma/biotech companies, CROs, and multiple regulatory agencies convened to actively discuss the most current topics of interest in bioanalysis. The 17th WRIB included 3 Main Workshops and 7 Specialized Workshops that together spanned 1 week to allow an exhaustive and thorough coverage of all major issues in bioanalysis of biomarkers, immunogenicity, gene therapy, cell therapy and vaccines. Moreover, in-depth workshops on "EU IVDR 2017/746 Implementation and impact for the Global Biomarker Community: How to Comply with these NEW Regulations" and on "US FDA/OSIS Remote Regulatory Assessments (RRAs)" were the special features of the 17th edition. As in previous years, WRIB continued to gather a wide diversity of international, industry opinion leaders and regulatory authority experts working on both small and large molecules as well as gene, cell therapies and vaccines to facilitate sharing and discussions focused on improving quality, increasing regulatory compliance, and achieving scientific excellence on bioanalytical issues. This 2023 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2023 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 3) covers the recommendations on Gene Therapy, Cell therapy, Vaccines and Biotherapeutics Immunogenicity. Part 1A (Mass Spectrometry Assays and Regulated Bioanalysis/BMV), P1B (Regulatory Inputs) and Part 2 (Biomarkers, IVD/CDx, LBA and Cell-Based Assays) are published in volume 16 of Bioanalysis, issues 8 and 9 (2024), respectively.

MSB11456 est un candidat biosimilaire du médicament biologique de référence tocilizumab. Sa similarité au tocilizumab de référence, homologué aux États-Unis (tocilizumab-EU) et approuvé dans l'Union Européenne (tocilizumab-UE), a été démontrée chez des volontaires sains pour la pharmacologie, la tolérance et l'immunogénicité. L'objectif était d'évaluer l'efficacité, la tolérance et l'immunogénicité de MSB11456 versus tocilizumab-UE chez des patients atteints de polyarthrite rhumatoïde (PR) active modérée à sévère, ayant présenté une réponse clinique inadéquate à au moins 1 DMARD et recevant une dose stable de méthotrexate. Les patients ont été randomisés pour recevoir en double aveugle des injections sous-cutanées de 162 mg de MSB11456 ou de tocilizumab-UE pendant 24 semaines (S). À S24, les patients recevant tocilizumab-UE ont été de nouveau randomisés pour poursuivre le traitement par tocilizumab-UE ou switcher vers un traitement par MSB11456 jusqu'à S52. Le critère principal d'efficacité - variation du score d'activité de la maladie DAS28-VS de l'inclusion à S24, a été évalué à l'aide d'une analyse de covariance visant à comparer les moyennes des moindres carrés (MMC) de MSB11456 et du tocilizumab-UE ; MSB11456 était considéré comme équivalent au tocilizumab-UE dès lors que l'intervalle de confiance (IC) à 95 % de la différence était compris entièrement dans l'intervalle d'équivalence prédéfini allant de -0,6 à 0,6. Les critères d'évaluation secondaires incluaient l'ACR20 à S24 et le DAS28-VS à S12. Les critères secondaires additionnels comprenaient notamment : les critères de réponse ACR50/70, la variation du DAS28-CRP, l'indice simplifié de l'activité de la maladie (SDAI), l'indice de l'activité clinique de la maladie (CDAI) à divers temps jusqu'à S52, ainsi que l'évaluation de l'immunogénicité et de la tolérance. Les résultats présentés concernent la période allant jusqu'à S24. Des diminutions cliniquement significatives des MMC du DAS28-VS ont été observées entre l'inclusion et S24 avec MSB11456 et le tocilizumab-UE (Tableau 1). L'IC à 95 % pour la différence des MMC de la variation du DAS28-VS entre l'inclusion et S24 étant entièrement compris dans l'intervalle d'équivalence prédéfini, l'équivalence thérapeutique de MSB11456 et du tocilizumab-UE a été démontrée. Toutes les analyses de sensibilité et les analyses des autres critères d'efficacité ont confirmé ce résultat. Les résultats d'immunogénicité ont été similaires entre les deux groupes de traitement. Les événements indésirables apparus sous traitement (EIAT) ont généralement été légers ou modérés et sont survenus à des fréquences similaires avec les deux médicaments. Aucune différence statistiquement significative n'a été observée tant sur la nature et la fréquence que sur tout autre caractéristique des EIAT associés au traitement ou des EIAT graves. L'équivalence d'efficacité, de tolérance et d'immunogénicité entre MSB11456 et tocilizumab de référence-UE a été démontrée à S24 chez des patients atteints de PR active modérée à sévère. Ces données soutiennent la similarité pharmacologique et clinique de ces deux médicaments biologiques ; par conséquent, MSB11456 peut être considéré comme un biosimilaire du tocilizumab-UE.

BACKGROUND:Tocilizumab, a recombinant monoclonal immunoglobulin G, targets the interleukin-6 receptor. MSB11456 is a proposed tocilizumab biosimilar. OBJECTIVES:To assess pharmacokinetic equivalence of intravenous MSB11456 to US-licensed tocilizumab. RESEARCH DESIGN AND METHODS:In this double-blind, parallel-group, single-dose study, 128 healthy adults were randomized to a single one-hour 8 mg/kg IV infusion of either MSB11456 or US-licensed tocilizumab. Blood samples were collected pre-dose and at regular intervals up to day 48 post-dose. The primary endpoint pharmacokinetic parameter was analyzed using analysis of variance (ANOVA) model on the natural logarithm of the endpoint (AUC0-last), with treatment as a fixed effect. Immunogenicity and safety data were summarized descriptively. RESULTS:Subjects received either MSB11456 (N = 62) or US-licensed tocilizumab (N = 66). Pharmacokinetic bioequivalence, defined as 90% confidence intervals for the geometric least squares mean ratio entirely contained within the 80.00% to 125.00% equivalence limits, was demonstrated between MSB11456 and US-licensed tocilizumab for the primary and secondary pharmacokinetic endpoints. Anti-drug antibody responses, frequency of neutralizing antibodies against tocilizumab, and safety profiles showed no notable between-treatment differences. Safety was comparable between treatments. CONCLUSIONS:Pharmacokinetic similarity of MSB11456 and US-licensed tocilizumab was demonstrated, with comparable immunogenicity and safety profiles, supporting MSB11455 as a biosimilar to US-licensed tocilizumab. The trial is registered at EudraCT, number 2019-003484-22.

Background MSB11456 is a biosimilar to currently marketed originator tocilizumab. It has demonstrated pharmacologic, safety and immunogenic similarity to the US-licensed and EU-approved tocilizumab in healthy subjects. Objectives To evaluate the efficacy, immunogenicity and safety of MSB11456 versus EU-approved tocilizumab in patients with moderate to severe rheumatoid arthritis (RA) who had experienced inadequate clinical response to ≥1 disease-modifying antirheumatic drug and were receiving a stable dose of methotrexate. Methods Patients were randomised to double-blind subcutaneous injections of 162 mg MSB11456 or EU-approved tocilizumab for 24 weeks (W). At W24, patients receiving EU-approved tocilizumab were re-randomized to continue their treatment or to switch to MSB11456 up to W52. Those receiving MSB11456 continued MSB11456 for the 52W. Primary efficacy endpoint - change from baseline in Disease Activity Score-28 Joint Count (DAS28)-erythrocyte sedimentation rate (ESR) at W24 - was analysed using analysis of covariance to determine the least squares mean (LSM) difference between MSB11456 and EU-approved tocilizumab; MSB11456 was considered equivalent to EU-approved tocilizumab if the 95% confidence interval (CI) for this difference was entirely within the equivalence interval -0.6 to 0.6. Secondary endpoints were 20% improvement in American College of Rheumatology core set measures (ACR20) at W24 and DAS28-ESR at W12. Additional endpoints included ACR50/70, change in DAS28-C-reactive protein (CRP), Simplified Disease Activity Index (SDAI), Clinical Disease Activity Index (CDAI) at various time points up to W52, and evaluation of immunogenicity and safety. Results up to W24 are presented. Results Clinically relevant LSM decreases from baseline in DAS28-ESR were observed at W24 with MSB11456 and EU-approved tocilizumab (Table 1). As the 95% CI for the LSM difference in the change from baseline in DAS28-ESR between treatments was fully included within the predefined equivalence interval, therapeutic equivalence of MSB11456 and EU-approved tocilizumab was demonstrated. All sensitivity and other efficacy endpoint analyses supported this finding. Immunogenicity results were similar in both treatment groups. Treatment-emergent adverse events (TEAEs) were usually mild or moderate and occurred at similar frequency with both drugs. There were no discernible patterns in terms of the nature, frequency or other characteristics of serious or treatment-related TEAEs to suggest a difference between drugs. Conclusion Equivalent efficacy, immunogenicity and safety at W24 of MSB11456 and EU-approved tocilizumab was demonstrated in patients with moderate to severe RA. Pharmacological and clinical similarity of these drugs was supported, therefore MSB11456 can be considered as biosimilar to EU-approved tocilizumab. Acknowledgements We thank the patients and the investigators of the working group. Disclosure of Interests Anna Zubrzycka-Sienkiewicz: None declared, Maria Misterska-Skora: None declared, Małgorzata Socik Pojawa: None declared, Kamilla Klama: None declared, Martin Ullmann Employee of: Fresenius Kabi SwissBioSim, Corinne Petit-Frere Employee of: Fresenius Kabi SwissBioSim, Andras Illes Employee of: Fresenius Kabi SwissBioSim, Emmanuelle Vincent Employee of: Fresenius Kabi SwissBioSim, Joëlle Monnet Employee of: Fresenius Kabi SwissBioSim, Jan Brzezicki: None declared.Efficacy endpoints – ITTMSB11456 (N=302)EU-approved tocilizumab (N=302)DAS28-ESR change from baseline at W24, LSM (SE)a-3.53 (0.11)-3.54 (0.11)LSM difference (95% CI)a0.01 (-0.16, 0.18)ACR20 at W24, n (%)244 (80.8)256 (84.8)Difference (95% CI)b-3.94 (-9.97, 2.11)DAS28-ESR change from baseline at W12, LSM (SE)a-3.13 (0.10)-3.12 (0.10)LSM difference (95% CI)a0.01 (-0.21, 0.19)DAS28-CRP change from baseline at W24, mean (standard deviation)-2.85 (1.22)-2.88 (1.11)ACR50 at W24, n (%)183 (60.6)188 (62.3)ACR70 at W24, n (%)118 (39.1)116 (38.4)DAS-ESR remission or LDA at W24, n/N (%)182/277 (65.7)191/285 (67.0)DAS-CRP responder at W24, n/N (%)46/276 (16.7)55/285 (19.3)CDAI remission or LDA at W24, n/N (%)172/278 (61.9)177/286 (61.9)SDAI remission or LDA at W24, n/N (%)174/276 (63.1)181/285 (63.5)aANCOVA (analysis of covariance) model with fixed effects for treatment and previous biologic for RA exposure and baseline DAS28-ESR as covariateb95% stratified Newcombe CI adjusting for previous biologic for RA exposureITT, intent to treat; LDA, low disease activity; SE, standard error

BACKGROUND:Tocilizumab is a monoclonal immunoglobulin G interleukin-6 receptor antagonist. MSB11456 is a proposed tocilizumab biosimilar. OBJECTIVE:To determine the pharmacokinetic equivalence of a single subcutaneous injection of MSB11456, when delivered via autoinjector (AI) and prefilled syringe (PFS), in healthy adult subjects. RESEARCH DESIGN AND METHODS:In this randomized, open-label, single fixed-dose, crossover study, 91 subjects received subcutaneous administration of tocilizumab 162 mg via AI and PFS presentations. The primary endpoint pharmacokinetic parameters were analyzed using analysis of variance. Safety data were summarized descriptively. RESULTS:There were no differences in pharmacokinetic parameters between presentations, and safety parameters were comparable. The 90% confidence intervals for the geometric least squares mean ratios of all primary pharmacokinetic parameters were contained within the predefined 80.00% to 125.00% bioequivalence limits, indicating pharmacokinetic equivalence between the AI and PFS. CONCLUSIONS:MSB11456 administration via AI was bioequivalent to administration via PFS. MSB11456 can be administered by AI or PFS, increasing the available range of self-injection devices. TRIAL REGISTRATION:The trial is registered at EudraCT, number 2020-003419-86.

The determination of a tailored anti-drug antibody (ADA) testing strategy is based on the immunogenicity risk assessment to allow a correlation of ADAs with changes to pharmacokinetics, efficacy, and safety. The clinical impact of ADA formation refines the immunogenicity risk assessment and defines appropriate risk mitigation strategies. Health agencies request for high-risk biotherapeutics to extend ADA monitoring for patients that developed an ADA response to the drug until ADAs return to baseline levels. However, there is no common understanding in which cases an extension of ADA follow-up sampling beyond the end of study (EOS) defined in the clinical study protocol is required. Here, the Immunogenicity Strategy Working Group of the European Immunogenicity Platform (EIP) provides recommendations on requirements for an extension of ADA follow-up sampling in clinical studies where there is a high risk of serious consequences from ADAs. The importance of ADA evaluation during a treatment-free period is recognized but the decision whether to extend ADA monitoring at a predefined EOS should be based on evaluation of ADA data in the context of corresponding clinical signals. If the clinical data set shows that safety consequences are minor, mitigated, or resolved, further ADA monitoring may not be required despite potentially detectable ADAs above baseline. Extended ADA monitoring should be centered on individual patient benefit.

Background Tocilizumab is a recombinant humanized monoclonal immunoglobulin G1 antibody against the interleukin-6 receptor (IL-6 R). MSB11456 is a proposed tocilizumab biosimilar. Objectives To assess the pharmacokinetic and pharmacodynamic similarity of MSB11456 to both US-licensed and EU-approved tocilizumab. Methods Healthy adult volunteers (N = 685) received a single 162 mg subcutaneous injection of MSB11456, US-licensed tocilizumab, or EU-approved tocilizumab in this randomized, double-blind, parallel-group study. Blood samples were taken pre-dose and for up to 48 days post-dose. Primary endpoint pharmacokinetic parameters were analyzed using analysis of covariance. Secondary pharmacodynamic measures included serum-soluble IL-6 R and serum C-reactive protein. Safety data were analyzed descriptively. Results Pharmacokinetic equivalence (with all corresponding 90% confidence intervals for the geometric least squares mean ratios within the predefined 80.00% to 125.00% equivalence margin) was demonstrated between MSB11456 and both US-licensed and EU-approved tocilizumab, as well as between the reference products. Pharmacodynamic analyses demonstrated similarity of MSB11456 and both US-licensed and EU-approved tocilizumab, as well as between the reference products. Safety, tolerability, and immunogenicity were comparable between treatments. Conclusion Pharmacokinetic and pharmacodynamic similarity of MSB11456, US-licensed tocilizumab, and EU-approved tocilizumab were demonstrated, and the three products had comparable immunogenicity and safety, supporting MSB11456 as a biosimilar to tocilizumab. PLAIN LANGUAGE SUMMARY Tocilizumab is a biologic drug that is used to treat autoimmune diseases, including rheumatoid arthritis. Biologic drugs are very important for the treatment of autoimmune diseases, but their costs limit accessibility. Therefore, the availability of biosimilars, which are biologics that are very similar in structure and function to an existing biologic drug, may provide a significant cost advantage for national healthcare programs and consumers. MSB11456 is a proposed tocilizumab biosimilar. Our study tested the pharmacokinetic and pharmacodynamic similarity of MSB11456 to the approved formulations of tocilizumab in the US and EU (US-licensed and EU-approved tocilizumab) in a large group of healthy adults. Volunteers received a single 162 mg subcutaneous injection of MSB11456, US-licensed tocilizumab, or EU-approved tocilizumab in this randomized, double-blind, parallel-group study. Blood samples were taken before and regularly after the injection, and safety was monitored. We showed that the pharmacokinetics and pharmacodynamics of MSB11456, US-licensed and EU-approved tocilizumab were sufficiently similar to claim equivalence between the three products. Safety and immunogenicity were also comparable between the three treatments. These findings suggest that MSB11456 can be considered as a biosimilar to tocilizumab. Biosimilars have improved price competition and led to a reduction in the net costs of biologics, so tocilizumab biosimilars can be expected to contribute to this and potentially improve access to the best available care.

Abstract MSB11455 is a proposed biosimilar to the currently licensed reference pegfilgrastim (Neulasta®). This study was designed primarily to compare the immunogenicity of MSB11455 and Neulasta®. As secondary objectives, the safety and tolerability of MSB11455 and Neulasta® were also compared. Healthy adult subjects were randomized to either MSB11455 or Neulasta®, stratified by antipolyethylene glycol (PEG) antibody status at screening and study site. Subjects received a single subcutaneous dose of MSB11455 or Neulasta® (both 6 mg/0.6 mL) on day 1 of each of two study periods (same product in both periods), separated by a washout of 28‐35 days. Immunogenicity samples were taken predose and up to day 84 post–first dose. Noninferiority was confirmed if the upper limit of the exact one‐sided adjusted 95% confidence interval (CI) for the difference in antidrug antibody (ADA)‐positive rates was < 10%. Safety was assessed throughout the study. Overall, 336 subjects were randomized and treated (N = 168 in each group). Noninferiority of MSB11455 over Neulasta® was demonstrated for immunogenicity; the difference in confirmed treatment‐induced ADA‐positive rate between MSB11455 and Neulasta® was −0.6% (upper limit of the exact one‐sided adjusted 95% CI: 6.25%). ADAs were mostly directed against the PEG moiety of pegfilgrastim. No filgrastim‐specific neutralizing antibodies were detected in either treatment group. Safety and tolerability were as expected for pegfilgrastim, and comparable between treatments. This study supports and strengthens the available evidence for the biosimilarity of MSB11455 to Neulasta®.

To compare the safety, efficacy, and immunogenicity of MSB11022 (acetate-buffered formulation), an adalimumab biosimilar, with the reference product. AURIEL-RA study was a phase 3, multicenter, randomized, double-blind, parallel group trial (NCT03052322). Patients with moderately-to-severely active rheumatoid arthritis (RA) with an inadequate response to methotrexate were randomized 1:1 to MSB11022 or reference adalimumab. The primary endpoint was the incidence of treatment-emergent adverse events of special interest (AESIs) (predefined as hypersensitivity) up to week 52. The key secondary endpoint was ACR20 (≥ 20% improvement in American College of Rheumatology core set measurements from baseline) at week 12. Other efficacy endpoints, quality of life, immunogenicity, and pharmacokinetic parameters were evaluated up to week 52. Secondary safety endpoints were evaluated up to week 52 and at a 4-month safety follow-up. In total, 288 patients were randomized. The proportion of patients experiencing ≥ 1 treatment-emergent AESI up to week 52 was similar between trial arms: 6 patients (4.2%; 95% CI 1.56, 8.91) receiving MSB11022, and 8 patients (5.5%; 95% CI 2.41, 10.58) receiving reference adalimumab. No clinically meaningful differences in efficacy, quality of life, or immunogenicity were seen between treatment arms up to week 52. No notable difference in the incidence of treatment-emergent adverse events was observed between treatment arms up to the end of the follow-up period. These results suggest MSB11022 and reference adalimumab are similar in patients with moderately-to-severely active rheumatoid arthritis in terms of safety, immunogenicity, and efficacy. AURIEL-RA provides evidence to support the similarity of MSB11022 and adalimumab.

MSB11022 is a proposed adalimumab biosimilar.

Background: Adalimumab is a fully human anti-TNF monoclonal antibody indicated for the treatment of multiple inflammatory disorders, including rheumatoid arthritis (RA). MSB11022 is a proposed adalimumab biosimilar that has been shown to be structurally and functionally similar to the adalimumab reference product1. MSB11022 has been developed in two formulations, in a citrate-based buffer, and in a modified buffer and stabiliser. MSB11022 demonstrated bioequivalence and comparable safety, tolerability and immunogenicity profiles to reference adalimumab (both in citrate formulations) in a study in healthy volunteers2. Subsequently MSB11022 was shown to be therapeutically equivalent to reference adalimumab (both citrate formulations) in terms of efficacy, safety and immunogenicity in psoriasis patients in the 52-week, Phase III, pivotal AURIEL-PsO study3. MSB11022 (modified formulation) demonstrated bioequivalence and a comparable safety and immunogenicity profile to MSB11022 (citrate formulation) in an additional healthy volunteer study (EMR200588-003)4. Objectives: To evaluate safety, immunogenicity and efficacy of MSB11022 (modified formulation) in patients with moderately to severe active rheumatoid arthritis compared to reference adalimumab up to 52 weeks. Methods: RA patients receiving methotrexate were randomised 1:1 to MSB11022 (modified formulation) or reference adalimumab (citrate formulation) in the double-blind, multicentre, phase III AURIEL-RA study (NCT03052322). Safety, efficacy and immunogenicity endpoints were assessed at scheduled visits up to week 52 using descriptive statistical methods only. Safety was the primary objective and the study was not powered to demonstrate equivalent efficacy. Results: 288 RA patients were randomised (MSB11022, n = 143; reference adalimumab, n = 145). Patient baseline characteristics were comparable between treatment groups. Few adverse events of special interest (AESIs) of hypersensitivity (primary endpoint) were reported during the study and the proportions were similar across treatment arms. Efficacy endpoints, including the key secondary endpoint of American College of Rheumatology criteria 20% (ACR20) response rate at week 12, were similar between the treatment arms. The safety profiles of patients receiving MSB11022 and reference adalimumab were also similar through to week 52. There were no clinically meaningful differences in the incidence of anti-drug antibodies (ADAs) and neutralising antibodies (NAbs) between treatment arms up to week 52. Key results are presented in Table 1. Conclusion: MSB11022 (modified formulation) and reference adalimumab had similar safety, immunogenicity and efficacy profiles over 52 weeks in patients with RA, supplementing the clinical data collected with MSB11022 (citrate formulation) in healthy volunteers and psoriasis patients References: [1] Magnenat L, et al. MAbs 2017;9(1):127-39. [2] Hyland E, et al. Br J Clin Pharmacol 2016;82(4):983-93. [3] Hercogova J, et al. J Am Acad Dermatol 2018;79(3):AB21. [4] Fresenius Kabi. Data on File 2019 Disclosure of Interests: Christopher Edwards Grant/research support from: Abbvie, BMS, Biogen, Celgene, Fresenius, Janssen, Lilly, Mundipharma, Pfizer, MSD, Novartis, Roche, Samsung, Sanofi, UCB, Consultant for: Abbvie, BMS, Biogen, Celgene, Fresenius, Janssen, Lilly, Mundipharma, Pfizer, MSD, Novartis, Roche, Samsung, Sanofi, UCB, Speakers bureau: Abbvie, BMS, Biogen, Celgene, Fresenius, Janssen, Lilly, Mundipharma, Pfizer, MSD, Novartis, Roche, Samsung, Sanofi, UCB, Veronica Chyrok Employee of: Former employee of Fresenius Kabi SwissBioSim , Joëlle Monnet Employee of: Employee of Fresenius Kabi SwissBioSim, Martin Ullmann Shareholder of: Amgen, BMS, Employee of: Employee of Fresenius Kabi SwissBioSim, Pantelis Vlachos: None declared

MSB11022 is a proposed adalimumab biosimilar that has shown structural and functional similarity to the reference product (Humira®). In addition, it has demonstrated bioequivalence in healthy volunteers and comparable efficacy, safety and immunogenicity in psoriasis patients. This analysis aims to show comparability of MSB11022 and reference adalimumab on QoL in psoriasis. In the double-blind, multicenter, pivotal efficacy and safety AURIEL-PsO study, patients were randomized 1:1 to MSB11022 or reference adalimumab. At Week 16, PASI 50 responders receiving reference product were re-randomized 1:1 to continue treatment or switch to MSB11022 until Week 52. QoL was assessed at scheduled visits using the Dermatology Life Quality Index (DLQI) and EuroQol 5-dimension 5-level (EQ-5D-5L) questionnaires. For the subpopulation of patients with psoriatic arthritis, the Health Assessment Questionnaire-Disability Index (HAQ-DI) and the Psoriatic Arthritis Joint Activity-Visual Analog Scale (PJA-VAS) were also used. The per-protocol analysis included 203 and 191 patients in the MSB11022 and reference adalimumab arms, respectively. Similarity of efficacy, safety and immunogenicity between treatment arms was demonstrated, including equivalent efficacy for the primary endpoint of PASI 75 at Week 16. Improvements from baseline to Week 16 for all QoL scores were also comparable. At Week 16, the least square mean difference (95% CI) for MSB11022 versus reference adalimumab was −0.12 (−0.83, 0.58) for DLQI, 0 (−0.02, 0.02) for EQ-5D-5L, −1.39 (−4.16, 1.38) for EQ-5D-5L-VAS, 0.05 (−0.15, 0.25) for HAQ-DI (n=43) and 2.05 (−12.4, 16.5) for PJA-VAS (n=43). Improvements in QoL scores to Week 52 were equally maintained in all treatment arms, including the 96 patients who switched from reference adalimumab to MSB11022 at Week 16. Treatment with MSB11022 and reference adalimumab resulted in comparable improvements in QoL at Week 16. QoL improvements were maintained to Week 52, and no impact of switching from reference product to MSB11022 was observed.

The American Association of Pharmaceutical Scientists (AAPS) biosimilar focus group on nonclinical and clinical assays has developed this manuscript to guide the industry on best practices and testing strategies when developing neutralizing antibody (NAb) assays for biosimilar programs. The immunogenicity assessment to biosimilar and originator drug products is one of the key aspects of clinical programs for biosimilars to demonstrate biosimilarity. Establishing that there are no clinically meaningful differences in immune response between a proposed product and the originator product is a key element in the demonstration of biosimilarity. It is critical to collect, evaluate, and compare the safety and immunogenicity data from the clinical pharmacology, safety, and/or efficacy studies especially when the originator drug product is known to have potential for immune-mediated toxicity. This manuscript aims to provide a comprehensive review and recommendations on assay formats, critical reagents, approaches to method development, and validation of the neutralizing antibody assays in extrapolation within the scope of biosimilar drug development programs. Even if there are multiple options on the development and validation of NAb assays for biosimilar programs, the type of drug and its MoA will help determine the assay format and technical platform for NAb assessment (e.g., cell-based or non-cell-based assay). We recommend to always perform a one-assay approach as it is better to confirm the biosimilarity using one-assay for NAb. If a one-assay approach is not feasible, then a two-assay format may be used. This manuscript will provide all the details necessary to develop NAb assays for biosimilars.

Background: Adalimumab is a fully human anti-TNF mAb, indicated for the treatment of multiple inflammatory disorders. MSB11022 is a proposed adalimumab biosimilar that has shown analytical similarity [1] and bioequivalence to US-licensed and EU-approved adalimumab originator, as well as comparable safety, tolerability and immunogenicity in a phase I trial [2]. Objectives: The aims of this multicentre, double-blind, parallel-group, 52-week phase III study (AURIEL-PsO, NCT02660580) were to demonstrate equivalence in efficacy (Psoriasis Area and Severity Index [PASI] 75) and to compare the safety and immunogenicity of MSB11022 vs. adalimumab originator in patients with moderate-to-severe chronic plaque psoriasis. This study was designed in-line with the biosimilar regulatory framework as part of the totality of evidence to confirm similarity and rationale for extrapolation. Methods: A total of 443 eligible patients (391 evaluable, including 43 with psoriatic arthritis) from 69 sites in 12 countries were randomised 1:1 and treated with MSB11022 (n=202) or adalimumab originator (n=189) (80 mg subcutaneously [SC] on day 1; 40 mg SC every other week from weeks 2−14). The primary endpoint was PASI 75 at week 16; equivalence was established if the 95% confidence interval (CI) for the treatment difference was within ±18%. Secondary endpoints included % change from baseline in PASI (equivalence confirmed if 95% CI within ±15%), Physician Global Assessment (PGA), quality of life (QoL), immunogenicity and safety. Interim results at week 16 are presented. Results: Patient baseline characteristics were comparable between MSB11022 and adalimumab originator groups: mean age 44.8 vs. 42.4 years, male 66.8% vs. 68.3%, mean PASI score 20.7 vs. 21.2, respectively. PASI 75 scores were 89.6% for MSB11022 and 91.5% for adalimumab originator (difference −1.9% [95% CI −7.82–4.16]). Mean % change from baseline in PASI was −90.6% for MSB11022 and −91.7% for adalimumab originator (difference −1.0% [95% CI −1.23–2.98]). PGA and QoL scores were comparable between treatment groups. The incidence of treatment-emergent adverse events (TEAEs)/serious TEAEs was 51.1/3.6% for MSB11022 and 53.2/2.7% for adalimumab originator. Immunogenicity profiles of MSB11022 and adalimumab originator were also similar and consistent. Conclusions: Week 16 results of this phase III confirmatory study demonstrated equivalent efficacy and similar safety and immunogenicity profiles for MSB11022 vs. adalimumab originator at 16 weeks in patients with moderate-to-severe chronic psoriasis. References [1] Magnenat L, et al. Demonstration of physicochemical and functional similarity between the proposed biosimilar adalimumab MSB11022 and Humira®. MAbs. 2017;9:127–139. [2] Hyland E, et al. Comparison of the pharmacokinetics, safety, and immunogenicity of MSB11022, a biosimilar of adalimumab, with Humira® in healthy subjects. Br J Clin Pharmacol. 2016;82:983–93. Disclosure of Interest: J. Hercogova Grant/research support from: Fresenius Kabi, Consultant for: Fresenius Kabi, K. Papp Grant/research support from: Fresenius Kabi, C. Edwards Grant/research support from: Fresenius Kabi, Consultant for: Fresenius Kabi, V. Chyrok Employee of: Fresenius Kabi, T. Halady Employee of: Fresenius Kabi, M. Ullmann Employee of: Fresenius Kabi, P. Vlachos Consultant for: Fresenius Kabi

The 2018 12th Workshop on Recent Issues in Bioanalysis took place in Philadelphia, PA, USA on April 9-13, 2018 with an attendance of over 900 representatives from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. WRIB was once again a 5-day full immersion in bioanalysis, biomarkers and immunogenicity. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small- and large-molecule bioanalysis involving LCMS, hybrid LBA/LCMS and LBA/cell-based assays approaches. This 2018 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2018 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 3) covers the recommendations for large molecule bioanalysis, biomarkers and immunogenicity using LBA and cell-based assays. Part 1 (LCMS for small molecules, peptides, oligonucleotides and small molecule biomarkers) and Part 2 (hybrid LBA/LCMS for biotherapeutics and regulatory agencies' inputs) are published in volume 10 of Bioanalysis, issues 22 and 23 (2018), respectively.