Increased potency of recombinant VWF D′D3 albumin fusion proteins engineered for enhanced affinity for coagulation factor VIII
| Autor: | Anton Zalewski, Arna Andrews, Isabelle Glauser, Marcel Mischnik, Matthew P. Hardy, Anne M Verhagen, Kerstin Emmrich, Sabine Pestel, Con Panousis, Philipp Claar, Victor Turnbull, Chao-Guang Chen, Michael J. Wilson, Gregory T. Bass, Elmar Raquet, Steve K. Dower, Jenny Chia, Thomas Weimer, Vesna Tomasetig |
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| Rok vydání: | 2021 |
| Předmět: |
congenital
hereditary and neonatal diseases and abnormalities Factor VIII biology Chemistry Recombinant Fusion Proteins Hematology Pharmacology Hemophilia A Fusion protein Recombinant Proteins In vitro Rats law.invention Directed mutagenesis Von Willebrand factor Coagulation law In vivo Albumins hemic and lymphatic diseases von Willebrand Factor biology.protein Recombinant DNA Animals Potency |
| Zdroj: | Journal of Thrombosis and Haemostasis. 19:2710-2725 |
| ISSN: | 1538-7836 |
| Popis: | BACKGROUND We have recently reported on a recombinant von Willebrand factor (VWF) D'D3 albumin fusion protein (rD'D3-FP) developed to extend the half-life of coagulation factor VIII (FVIII) for the treatment of hemophilia A. Based on predictive modelling presented in this study, we hypothesized that modifying rD'D3-FP to improve FVIII interaction would reduce exchange with endogenous VWF and provide additional FVIII half-life benefit. OBJECTIVES The aim of this study was to identify novel rD'D3-FP variants with enhanced therapeutic efficacy in extending FVIII half-life. METHODS Through both directed mutagenesis and random mutagenesis using a novel mammalian display platform, we identified novel rD'D3-FP variants with increased affinity for FVIII (rVIII-SingleChain) under both neutral and acidic conditions and assessed their ability to extend FVIII half-life in vitro and in vivo. RESULTS In rat preclinical studies, rD'D3-FP variants with increased affinity for FVIII displayed enhanced potency, with reduced dose levels required to achieve equivalent rVIII-SingleChain half-life extension. In cell-based imaging studies in vitro, we also demonstrated reduced dissociation of rVIII-SingleChain from the rD'D3-FP variants within acidic endosomes and more efficient co-recycling of the rD'D3-FP/rVIII-SingleChain complex via the FcRn recycling system. CONCLUSIONS In summary, at potential clinical doses, the rD'D3-FP variants provide marked benefits with respect to dose levels and half-life extension of co-administered FVIII, supporting their development for use in the treatment of hemophilia A. |
| Databáze: | OpenAIRE |
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