title: A sensitive venous bleeding model in haemophilia A mice: effects of two recombinant FVIII products (N8 and Advate(®)).
abstract: Haemostatic effect of compounds for treating haemophilia can be evaluated in various bleeding models in haemophilic mice. However, the doses of factor VIII (FVIII) for normalizing bleeding used in some of these models are reported to be relatively high. The aim of this study was to establish a sensitive venous bleeding model in FVIII knock out (F8-KO) mice, with the ability to detect effect on bleeding at low plasma FVIII concentrations. We studied the effect of two recombinant FVIII products, N8 and Advate(®), after injury to the saphenous vein. We found that F8-KO mice treated with increasing doses of either N8 or Advate(®) showed a dose-dependent increase in the number of clot formations and a reduction in both average and maximum bleeding time, as well as in average blood loss. For both compounds, significant effect was found at doses as low as 5 IU kg(-1) when compared with vehicle-treated F8-KO mice. Normalization of maximum bleeding time was found at doses equal to or above 10 IU kg(-1) N8 or Advate(®), corresponding to plasma concentrations of approximately 10% of the level in wild type mice. The present study adds a new model to the armamentarium of bleeding models used for evaluation of pro-coagulant compounds for treatment of haemophilia. Interestingly, the vena saphena model proved to be sensitive towards FVIII in plasma levels that approach the levels preventing bleeding in haemophilia patients, and may, thus, in particular be valuable for testing of new long-acting variants of e.g. FVIII that are intended for prophylaxis.
authors.affiliation: Department of Disease Biology, Faculty of Life Science, University of Copenhagen, Copenhagen, Denmark. aekr@novonordisk.com
authors.forename: A E
authors.initials: AE
authors.lastname: Pastoft
authors.forename: J
authors.initials: J
authors.lastname: Lykkesfeldt
authors.forename: M
authors.initials: M
authors.lastname: Ezban
authors.forename: M
authors.initials: M
authors.lastname: Tranholm
authors.forename: H C
authors.initials: HC
authors.lastname: Whinna
authors.forename: B
authors.initials: B
authors.lastname: Lauritzen
mesh_terms: D000818:Animals; D001760:Bleeding Time; D004195:Disease Models, Animal; D005169:Factor VIII / Q000008:administration & dosage*; D005260:Female; D006467:Hemophilia A / Q000150:complications* / Q000235:genetics / Q000628:therapy*; D006470:Hemorrhage / Q000209:etiology* / Q000517:prevention & control / Q000628:therapy*; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011994:Recombinant Proteins / Q000008:administration & dosage; D012501:Saphenous Vein / Q000293:injuries
chemical_list: D011994:Recombinant Proteins; C577506:recombinant factor VIII N8; D005169:Factor VIII