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In vitro characterization of MOD ‐5014, a novel long‐acting carboxy‐terminal peptide (CTP)‐modified activated FVII
Author(s) -
BarIlan A.,
Livnat T.,
Hoffmann M.,
Binder L.,
Zakar M.,
Guy R.,
Felikman Y.,
Moschcovich L.,
Shenkman B.,
Monroe D.,
Hershkovitz O.,
Kenet G.,
Hart G.
Publication year - 2018
Publication title -
haemophilia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.213
H-Index - 92
eISSN - 1365-2516
pISSN - 1351-8216
DOI - 10.1111/hae.13428
Subject(s) - in vitro , peptide , in vivo , antithrombin , medicine , recombinant dna , thrombin , heparin , pharmacology , biochemistry , chemistry , microbiology and biotechnology , platelet , immunology , biology , gene
Introduction Recombinant FVII a ( rFVII a) is an effective treatment for haemophilia through frequent administration. However, the short half‐life of rFVII a decreases its prophylactic ability to reduce bleeding. Carboxy‐terminal peptide ( CTP )‐modified FVII a ( MOD ‐5014) is a long‐acting rFVII a developed for on‐demand treatment of haemophilia using either an intravenous or subcutaneous injection with the aim of less frequent administrations, as well as for prophylactic use. Aim The comprehensive evaluation of the activity MOD ‐5014 vs commercially available rh FVII a, as well as their interaction with cofactors and inhibitors. Methods The in vitro characterization included clotting activity, affinity by surface plasmon resonance, cleavage of synthetic substrates, thrombin generation ( TG ) and rotation thromboelastometry. Results Reduced specific activity was obtained for MOD ‐5014 compared to rh FVII a, while both compounds demonstrated comparable affinity to tissue factor ( TF ). MOD ‐5014 showed reduced TG when spiked at a similar concentration as rh FVII a, suggesting that an increased concentration might be needed in a clinical setting to provide initial haemostatic effect. MOD ‐5014 demonstrated a slightly lower affinity for binding to activated platelets and slightly lower proteolytic activity on the platelet surface, possibly as the fusion of CTP has the potential to sterically hinder binding to both the platelet membrane and to protein substrates. Both compounds showed a similar dose‐dependent stimulatory effect on clot formation, and both showed a similar deactivation pattern following incubation with TF pathway inhibitor ( TFPI ), antithrombin and heparin. Conclusion The comparable in vitro activity of MOD ‐5014 and rh FVII a paves the way for in vivo pharmacology evaluations of MOD ‐5014 in preparation for clinical studies.