Systematic Verification of Bioanalytical Similarity Between a Biosimilar and a Reference Biotherapeutic: Committee Recommendations for the Development and Validation of a Single Ligand-Binding Assay to Support Pharmacokinetic Assessments
Marini J.C.,Janssen Research and Development LLC |
Anderson M.,BDS Immunoassay Services |
Cai X.-Y.,Merck Ltd. |
Chappell J.,CPR Pharma Services Pty Ltd |
And 7 more authors.
Botanical Review | Year: 2014
For biosimilar drug development, it is critical to demonstrate similar physiochemical characteristics, efficacy, and safety of the biosimilar product compared to the reference product. Therefore, pharmacokinetic (PK) and immunogenicity (antidrug antibody, ADA) assays that allow for the demonstration of biosimilarity are critical. Under the auspices of the American Association of Pharmaceutical Scientists (AAPS) Ligand-Binding Assay Bioanalytical Focus Group (LBABFG), a Biosimilars Action Program Committee (APC) was formed in 2011. The goals of this Biosimilars APC were to provide a forum for in-depth discussions on issues surrounding the development and validation of PK and immunogenicity assays in support of biosimilar drug development and to make recommendations thereof. The Biosimilars APC’s recommendations for the development and validation of ligand-binding assays (LBAs) to support the PK assessments for biosimilar drug development are presented here. Analytical recommendations for the development and validation of LBAs to support immunogenicity assessments will be the subject of a separate white paper. © 2014, American Association of Pharmaceutical Scientists.
Voss M.,Ludwig Maximilians University of Munich |
Kunzel U.,Ludwig Maximilians University of Munich |
Kunzel U.,University of Oxford |
Higel F.,Sandoz Biopharmaceuticals Hexal AG |
And 20 more authors.
EMBO Journal | Year: 2014
Protein N-glycosylation is involved in a variety of physiological and pathophysiological processes such as autoimmunity, tumour progression and metastasis. Signal peptide peptidase-like 3 (SPPL3) is an intramembrane-cleaving aspartyl protease of the GxGD type. Its physiological function, however, has remained enigmatic, since presently no physiological substrates have been identified. We demonstrate that SPPL3 alters the pattern of cellular N-glycosylation by triggering the proteolytic release of active site-containing ectodomains of glycosidases and glycosyltransferases such as N-acetylglucosaminyltransferase V, β-1,3 N-acetylglucosaminyltransferase 1 and β-1,4 galactosyltransferase 1. Cleavage of these enzymes leads to a reduction in their cellular activity. In line with that, reduced expression of SPPL3 results in a hyperglycosylation phenotype, whereas elevated SPPL3 expression causes hypoglycosylation. Thus, SPPL3 plays a central role in an evolutionary highly conserved post-translational process in eukaryotes. Synopsis SPPL3 is a highly conserved eukaryotic intramembrane-cleaving GxGD-type aspartyl protease of undefined function. We show that SPPL3 liberates medial/trans-Golgi glycosyltransferases from their N-terminal membrane anchors to regulate the intracellular pool of active Golgi glycosyltransferases and the extent of N-glycan decoration of cellular glycoproteins. Loss of SPPL3 in vitro and in vivo is associated with more extensive N-glycosylation. Overexpression of active SPPL3, but not of an inactive mutant, leads to less extensive N-glycosylation. Constitutive secretion of Golgi glycosyltransferases such as GnT-V, β3GnT1 and β4GalT1 is dependent on cellular SPPL3 activity. SPPL3-dependent GnT-V endoproteolysis occurs close to GnT-V's predicted transmembrane domain. Changes in SPPL3 expression strongly affect intracellular glycosyltransferase levels, explaining the observed alterations in N-glycan composition. The intramembrane-cleaving GxGD-type aspartyl protease, SPPL3, controls the proteolytic release of the ectodomain of glycosyltransferases and glycosidases to regulate cellular N-glycosylation. © 2014 The Authors.
Da Silva A.,Sandoz Biopharmaceuticals Hexal AG |
Kronthaler U.,Sandoz Biopharmaceuticals Hexal AG |
Koppenburg V.,Sandoz Biopharmaceuticals Hexal AG |
Fink M.,Novartis |
And 5 more authors.
Leukemia and Lymphoma | Year: 2014
Biosimilar development involves a target-directed iterative process to ensure a similar product to the originator. Here we report the preclinical development of the proposed biosimilar rituximab (GP2013). Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. Pharmacological comparability between GP2013 and originator rituximab were confirmed in preclinical studies using clinical scale drug product. Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. This preclinical comparability exercise confirms that GP2013 and originator rituximab are pharmacologically similar. © 2014 Informa UK, Ltd.