title: A novel B-domain O-glycoPEGylated FVIII (N8-GP) demonstrates full efficacy and prolonged effect in hemophilic mice models.
abstract: Frequent infusions of intravenous factor VIII (FVIII) are required to prevent bleeding associated with hemophilia A. To reduce the treatment burden, recombinant FVIII with a longer half-life was developed without changing the protein structure. FVIII-polyethylene glycol (PEG) conjugates were prepared using an enzymatic process coupling PEG (ranging from 10 to 80 kDa) selectively to a unique O-linked glycan in the FVIII B-domain. Binding to von Willebrand factor (VWF) was maintained for all conjugates. Upon cleavage by thrombin, the B-domain and the associated PEG were released, generating activated FVIII (FVIIIa) with the same primary structure and specific activity as native FVIIIa. In both FVIII- and VWF-deficient mice, the half-life was found to increase with the size of PEG. In vivo potency and efficacy of FVIII conjugated with a 40-kDa PEG (N8-GP) and unmodified FVIII were not different. N8-GP had a longer duration of effect in FVIII-deficient mouse models, approximately a twofold prolonged half-life in mice, rabbits, and cynomolgus monkeys; however, the prolongation was less pronounced in rats. Binding capacity of N8-GP on human monocyte-derived dendritic cells was reduced compared with unmodified FVIII, resulting in several-fold reduced cellular uptake. In conclusion, N8-GP has the potential to offer efficacious prevention and treatment of bleeds in hemophilia A at reduced dosing frequency.
authors: Novo Nordisk, Måløv, Denmark. hrse@novonordisk.com
authors: Henning R
authors: HR
authors: Stennicke
authors: Marianne
authors: M
authors: Kjalke
authors: Ditte M
authors: DM
authors: Karpf
authors: Kristoffer W
authors: KW
authors: Balling
authors: Peter B
authors: PB
authors: Johansen
authors: Torben
authors: T
authors: Elm
authors: Kristine
authors: K
authors: Øvlisen
authors: Flemming
authors: F
authors: Möller
authors: Heidi L
authors: HL
authors: Holmberg
authors: Charlotte N
authors: CN
authors: Gudme
authors: Egon
authors: E
authors: Persson
authors: Ida
authors: I
authors: Hilden
authors: Hermann
authors: H
authors: Pelzer
authors: Henrik
authors: H
authors: Rahbek-Nielsen
authors: Christina
authors: C
authors: Jespersgaard
authors: Are
authors: A
authors: Bogsnes
authors: Anette A
authors: AA
authors: Pedersen
authors: Anne K
authors: AK
authors: Kristensen
authors: Bernd
authors: B
authors: Peschke
authors: Wendy
authors: W
authors: Kappers
authors: Frederik
authors: F
authors: Rode
authors: Lars
authors: L
authors: Thim
authors: Mikael
authors: M
authors: Tranholm
authors: Mirella
authors: M
authors: Ezban
authors: Eva H N
authors: EH
authors: Olsen
authors: Søren E
authors: SE
authors: Bjørn
mesh_terms: D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D004195:Disease Models, Animal; D005169:Factor VIII / Q000008:administration & dosage / Q000378:metabolism / Q000627:therapeutic use*; D005260:Female; D006031:Glycosylation; D006467:Hemophilia A / Q000188:drug therapy* / Q000473:pathology; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008954:Models, Biological; D011092:Polyethylene Glycols / Q000008:administration & dosage / Q000378:metabolism / Q000627:therapeutic use*; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D016896:Treatment Outcome
chemical_list: C579325:N8-GP compound; D011092:Polyethylene Glycols; D005169:Factor VIII