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Thromb Haemost 2010; 104(02): 243-251
DOI: 10.1160/TH09-10-0723
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Investigating clearance mechanisms for recombinant activated factor VII in a perfused liver model

Rupa S. Appa
1   Exploratory ADME, Biopharmaceutical Research Unit, Novo Nordisk A/S, Måløv, Denmark
,
Charlotte Theill
1   Exploratory ADME, Biopharmaceutical Research Unit, Novo Nordisk A/S, Måløv, Denmark
,
Lene Hansen
1   Exploratory ADME, Biopharmaceutical Research Unit, Novo Nordisk A/S, Måløv, Denmark
,
Judi Møss
2   M&S Hematology and Clinical Pharmacology, Novo Nordisk A/S, Bagsværd, Denmark
,
Carsten Behrens
3   Biopharm Chemistry, Novo Nordisk A/S, Måløv, Denmark
,
Else Marie Nicolaisen
4   NovoSeven® Research, Novo Nordisk A/S, Måløv, Denmark
,
Niels K. Klausen
5   Protein Chemistry, Biopharm, CMC Analysis and Formulation, Novo Nordisk A/S, Gentofte, Denmark
,
Michael S. Christensen
6   DMPK and Bioanalysis Preclinical Development, Novo Nordisk A/S, Måløv, Denmark
› Institutsangaben Financial support: This study was supported by Novo Nordisk A/S, Måløv, Denmark.
Weitere Informationen

Publikationsverlauf

Received: 23. Oktober 2009

Accepted after major revision: 29. März 2010

Publikationsdatum:
24. November 2017 (online)

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Summary

Clearance mechanisms for recombinant activated human FVII (rFVIIa; NovoSeven®), a heterogeneously glycosylated protein, have yet to be fully elucidated, but may involve the liver. The effects of the γ-carboxy glutamic acid (Gla) domain and the sialic acid content of the protein on rFVIIa clearance were investigated following intravenous administration of rFVIIa lacking the Gla domain, des(1–44) rFVIIa and asialo-rFVIIa in pharmacokinetic (PK) studies and perfused rat livers. PK parameters for both rFVIIa and des(1–44) rFVIIa had similar biphasic clearance profiles, as well as half-lives ([t½]=80 and 88 minutes, respectively), while asialo-rFVIIa was cleared quickly (t½=21 minutes) with a linear clearance profile. Perfused liver studies with all proteins (10 nM) mirrored the trends in profiles observed in the PK study. rFVIIa and des(1–44) rFVIIa were cleared to a similar extent, 41% and 35%, respectively, after 1 h, whereas plasma-derived FVII from humans (which has a higher sialylation content than rFVIIa) was cleared to a lesser extent (21%). Asialo-rFVIIa, on the other hand, was almost totally cleared and when an excess of asialo-orosomucoid was added to the perfusate, its clearance was significantly reduced (by 34%) and also for rFVIIa, albeit to a lesser extent (by 14%). Together these data suggest that carbohydrate receptor(s) (e.g. the asialoglycoprotein receptor, ASGPR) play a role in asialo-rFVIIa and rFVIIa clearance. In vivo and liver clearance data correlated well showing similar trends and indi-cated that rFVIIa clearance is not affected by the Gla domain, but rather by a subpopulation of N-glycosylated structures on rFVIIa.

Keywords

Recombinant activated factor VII (rFVIIa) - in situ rat liver perfusion - asialoglycoprotein receptor (ASGPR) - asialo-rFVIIa - asialo-orosomu-coid
 

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