Analytical similarity assessment of rituximab biosimilar CT-P10 to reference medicinal product

Abstract

CT-P10 (Truxima™) was recently approved as the world's first rituximab biosimilar product in the European Union (EU) and South Korea. To demonstrate biosimilarity of CT-P10 with the reference medicinal product (RMP), extensive 3-way similarity assessment has been conducted between CT-P10, EU-Rituximab and US-Rituximab, focusing on the physicochemical and biological quality attributes. A multitude of state-of-the-art analyses revealed that CT-P10 has identical primary and higher order structures compared to the original product. Purity/impurity profiles of CT-P10 measured by the levels of aggregates, fragments, non-glycosylated form and process-related impurities were also found to be comparable with those of RMPs. In terms of the post-translational modification, CT-P10 contains slightly less N-terminal pyro-glutamate variant, which has been known not to affect product efficacy or safety. Oligosaccharide profiling has revealed that, although CT-P10 contains the same conserved glycan species and relative proportion with the RMPs, the content of total afucosylated glycan in CT-P10 was slightly higher than in EU- or US-Rituximab. Nevertheless, the effect of the observed level of afucosylation in CT-P10 drug product on Fc receptor binding affinity or antibody-dependent cell-mediated cytotoxicity was found to be negligible based on the spiking study with highly afucosylated sample. Arrays of biological assays representative of known and putative mechanisms of action for rituximab have shown that biological activities of CT-P10 are within the quality range of RMPs. Recent results of clinical studies have further confirmed that the CT-P10 exhibits equivalent clinical efficacy and safety profiles compared to EU- and US-Rituximab. The current 3-way similarity assessment together with clinical study results confidently demonstrate that CT-P10 is highly similar with EU- and US-Rituximab in terms of physicochemical properties, biological activities, efficacy, and safety for its final approval as a biosimilar product.

On the Aggregation and Nucleation Mechanism of the Monoclonal Antibody Anti-CD20 Near Liquid-Liquid Phase Separation (LLPS)

“…7, initial conditions) corresponds to the expected spectra for Anti-CD20, as it shows the typical shape of a monoclonal antibody with a minimum around 220 nm and a maximum around 200 nm, suggesting a secondary structure dominated by β-sheet motif 54 . Moreover, these spectra correspond to the CD-spectra of Anti-CD20 from literature 55 .…”

“…Nonetheless, high concentration behaviour can differ from that observed at dilute conditions, since "crowding" effects, i.e. the increased chance of molecular interactions, deviation from thermodynamic ideality and higher-order interactions (e.g., B 23 , B 222 ) can significantly alter the net inter-protein interactions 55,61 . The contribution of these effects to aggregation propensity of structurally complex molecules such as mAbs, cannot be ruled out.…”

Effects of Glycan Structure on the Stability and Receptor Binding of an IgG4-Fc

“…18 Several analytical approaches have been developed to qualitatively and quantitatively monitor oligosaccharide profiles of mAbs, including fluorescence detection of 2-aminobenzamide labeled glycans and HPLC-MS analysis of released glycans, glycopeptides, and glycoproteins. 19 Analytical results suggest that variations of glycan structures often exist among biosimilars and reference products. 20 Alterations in glycan structures have been shown to affect physicochemical properties 11,12,21,22 and biological effector functions 13,17 of IgG1 or the crystallizable fragment of IgG1 (IgG1-Fc), but such knowledge is limited with regard to IgG4.…”

Anticorps monoclonaux biosimilaires

“…Cette approche également connue sous le nom de cartographie peptidique a aujourd'hui presque totalement remplacé les méthodes anciennes de séquençage des protéines, longues, fastidieuses et coûteuses, comme la dégradation d'Edman. Ainsi, des cartes peptidiques ont été réalisées pour deux AcM ou une protéine de fusion de référence et leurs biosimilaires (rituximab [12,13], adalimumab [14] et étanercept [15]). Dans ces études de comparabilité, la séquence d'acides aminés a été validée pour les biosimilaires du rituximab et de l'adalimumab, avec une couverture de séquence très proche ou égale à 100 %.…”

“…La structure quaternaire de l'anticorps (hétérodimère de 2 chaînes lourdes et légères) peut être confirmée par exemple par spectrométrie de masse. [12]. A contrario, les biosimilaires de l'infliximab ont montré des différences de comportement in vitro, notamment en ce qui concerne leur fixation au RFcγIIIa [19].…”