Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains.

Autor: Siegemund M; Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany., Oak P; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Hansbauer EM; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Allersdorfer A; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Utschick K; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Winter A; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Grasmüller C; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Galler G; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Mayer JP; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Weiche B; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Prassler J; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany., Kontermann RE; Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.; Stuttgart Research Center Systems Biology, University of Stuttgart, Stuttgart, Germany., Rothe C; Pieris Pharmaceuticals GmbH, Hallbergmoos, Germany.
Jazyk: angličtina
Zdroj: Frontiers in pharmacology [Front Pharmacol] 2021 Oct 25; Vol. 12, pp. 759337. Date of Electronic Publication: 2021 Oct 25 (Print Publication: 2021).
DOI: 10.3389/fphar.2021.759337
Abstrakt: Anticalin ® proteins have been proven as versatile clinical stage biotherapeutics. Due to their small size (∼20 kDa), they harbor a short intrinsic plasma half-life which can be extended, e.g., by fusion with IgG or Fc. However, for antagonism of co-immunostimulatory Tumor Necrosis Factor Receptor Superfamily (TNFRSF) members in therapy of autoimmune and inflammatory diseases, a monovalent, pharmacokinetically optimized Anticalin protein format that avoids receptor clustering and therefore potential activation is favored. We investigated the suitability of an affinity-improved streptococcal Albumin-Binding Domain (ABD) and the engineered Fab-selective Immunoglobulin-Binding Domain (IgBD) SpGC3Fab for plasma Half-Life Extension (HLE) of an OX40-specific Anticalin and bispecific Duocalin proteins, neutralizing OX40 and a second co-immunostimulatory TNFRSF member. The higher affinity of ABD fusion proteins to human serum albumin (HSA) and Mouse Serum Albumin (MSA), with a 4 to 5-order of magnitude lower K D compared with the binding affinity of IgBD fusions to human/mouse IgG, translated into longer terminal plasma half-lives ( t 1/2 ). Hence, the anti-OX40 Anticalin-ABD protein reached t 1/2 values of ∼40 h in wild-type mice and 110 h in hSA/hFcRn double humanized mice, in contrast to ∼7 h observed for anti-OX40 Anticalin-IgBD in wild-type mice. The pharmacokinetics of an anti-OX40 Anticalin-Fc fusion protein was the longest in both models ( t 1/2 of 130 h and 146 h, respectively). Protein formats composed of two ABDs or IgBDs instead of one single HLE domain clearly showed longer presence in the circulation. Importantly, Anticalin-ABD and -IgBD fusions showed OX40 receptor binding and functional competition with OX40L-induced cellular reactivity in the presence of albumin or IgG, respectively. Our results suggest that fusion to ABD or IgBD can be a versatile platform to tune the plasma half-life of Anticalin proteins in response to therapeutic needs.
Competing Interests: Authors AA, GG and BW were employed by Pieris Pharmaceuticals GmbH. Authors MS, PO, E-MH, KU, AW, CG, J-PM, JP and CR are employed by Pieris Pharmaceuticals GmbH. The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
(Copyright © 2021 Siegemund, Oak, Hansbauer, Allersdorfer, Utschick, Winter, Grasmüller, Galler, Mayer, Weiche, Prassler, Kontermann and Rothe.)
Databáze: MEDLINE
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