Biodistribution of the recombinant fusion protein linking coagulation factor IX with albumin (rIX-FP) in rats.

Introduction: The recombinant fusion protein linking coagulation factor IX with albumin (rIX-FP) is undergoing clinical trials for prophylaxis and on-demand treatment of haemophilia B patients. The aim of this study was to investigate the pharmacokinetics, whole-body and knee joint distribution of rIX-FP following intravenous administration to rats, compared with a marketed, non-fused rFIX and recombinant human albumin. Material and Methods: [H-3]-rIX-FP, [H-3]-rFIX or [H-3]-albumin were administered to rats followed by quantitative whole-body autoradiography over 24 or 240 hours, and the tissue distribution as well as elimination of radioactivity were measured. Results: Elimination of all radioactivity derived from the three proteins was shown to occur primarily via the urine. The tissue distribution of [H-3]-rIX-FP and [H-3]-rFIX (but not of [H-3]-albumin) was comparable, both penetrating predominantly into bone, and well-perfused tissues, suggesting that the rIX moiety determines the distribution pattern of rIX-FP, while the albumin moity is responsible for the prolonged plasma and tissue retention. Detailed knee-joint analysis indicated rapid presence of [H-3]-rIX-FP and [H-3]-rFIX in synovial and mineralised bone tissue, mostly localised to the zone of calcified cartilage. Longest retention times were observed in the bone marrow and the endosteum of long bones. Intriguingly, [H-3]-rIX-FP- and [H-3]-albumin-derived radioactive signals were detectable up to 240 hours, while [H-3]-rFIX-derived radioactivity rapidly declined after 1 hour post-dosing correlating to the extended plasma half-life of [H-3]-rIX-FP. Conclusion: The prolonged plasma and tissue retention of rIX-FP achieved by albumin fusion may allow a reduction in dosing frequency leading to increased therapeutic compliance and convenience. (C) 2014 The Authors. Published by Elsevier Ltd.