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

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...

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Autores principales: Siegemund, Martin, Oak, Prajakta, Hansbauer, Eva-Maria, Allersdorfer, Andrea, Utschick, Karoline, Winter, Alexandra, Grasmüller, Christina, Galler, Gunther, Mayer, Jan-Peter, Weiche, Benjamin, Prassler, Josef, Kontermann, Roland E., Rothe, Christine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8573339/
https://www.ncbi.nlm.nih.gov/pubmed/34759826
http://dx.doi.org/10.3389/fphar.2021.759337
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author Siegemund, Martin
Oak, Prajakta
Hansbauer, Eva-Maria
Allersdorfer, Andrea
Utschick, Karoline
Winter, Alexandra
Grasmüller, Christina
Galler, Gunther
Mayer, Jan-Peter
Weiche, Benjamin
Prassler, Josef
Kontermann, Roland E.
Rothe, Christine
author_facet Siegemund, Martin
Oak, Prajakta
Hansbauer, Eva-Maria
Allersdorfer, Andrea
Utschick, Karoline
Winter, Alexandra
Grasmüller, Christina
Galler, Gunther
Mayer, Jan-Peter
Weiche, Benjamin
Prassler, Josef
Kontermann, Roland E.
Rothe, Christine
author_sort Siegemund, Martin
collection PubMed
description 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.
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spelling pubmed-85733392021-11-09 Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains Siegemund, Martin Oak, Prajakta Hansbauer, Eva-Maria Allersdorfer, Andrea Utschick, Karoline Winter, Alexandra Grasmüller, Christina Galler, Gunther Mayer, Jan-Peter Weiche, Benjamin Prassler, Josef Kontermann, Roland E. Rothe, Christine Front Pharmacol Pharmacology 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. Frontiers Media S.A. 2021-10-25 /pmc/articles/PMC8573339/ /pubmed/34759826 http://dx.doi.org/10.3389/fphar.2021.759337 Text en Copyright © 2021 Siegemund, Oak, Hansbauer, Allersdorfer, Utschick, Winter, Grasmüller, Galler, Mayer, Weiche, Prassler, Kontermann and Rothe. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Siegemund, Martin
Oak, Prajakta
Hansbauer, Eva-Maria
Allersdorfer, Andrea
Utschick, Karoline
Winter, Alexandra
Grasmüller, Christina
Galler, Gunther
Mayer, Jan-Peter
Weiche, Benjamin
Prassler, Josef
Kontermann, Roland E.
Rothe, Christine
Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains
title Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains
title_full Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains
title_fullStr Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains
title_full_unstemmed Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains
title_short Pharmacokinetic Engineering of OX40-Blocking Anticalin Proteins Using Monomeric Plasma Half-Life Extension Domains
title_sort pharmacokinetic engineering of ox40-blocking anticalin proteins using monomeric plasma half-life extension domains
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8573339/
https://www.ncbi.nlm.nih.gov/pubmed/34759826
http://dx.doi.org/10.3389/fphar.2021.759337
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