Immunogenicity of novel recombinant human activated factor VII analogues on factor VII neonatally-tolerized rats

Summary

Recombinant activated factorVII (rFVIIa; NovoSeven®) has been widely used to treat bleeding in patients with haemophilia with inhibitors. To increase the intrinsic activity, analogues of rFVIIa (rFVIIa_Q, rFVIIaDV_Q, and rFVIIaDV_QA) with altered amino acid sequence at or near the active centre have been developed. The immunogenicity of these analogues was tested in a rat immune tolerance model.Neonatal rats received rFVIIa intraperitoneally on post-natal Day 1 and were subsequently challenged with rFVIIa in Freunds Incomplete Adjuvant subcutaneously on Days 10 and 24. Rats were tested for tolerance on Day 32; the tolerant cohort and a parallel cohort of untreated control rats were challenged with rFVIIa, rFVIIa_Q, rFVIIaDV_Q, or rFVIIaDV_QA on Days 46 and 76. Immune responses determined by enzymelinked immunosorbent assay (ELISA) on Day 84 showed no statistically significant difference between the responses in the four control cohorts. Immune responses were higher in the control than in the tolerant cohort. Compared with rFVIIa (4/16), there was no difference in the proportion of rats that broke tolerance following challenge with rFVIIaDV_Q (3/16) and rFVIIaDV_QA (7/16), whereas a statistically significant greater proportion broke tolerance after challenge with rFVIIa_Q (11/16). Therefore, in this model rFVIIaDV_Q or rFVIIa DVQA were not more immunogenic than rFVIIa.

Keywords

Immunogenicity - Neonatal rat model - rFVIIa - rFVIIa analogues - tolerance

• References

• 1 Abshire T, Kenet G. Recombinant factor VIIa: review of efficacy, dosing regimens and safety in patients with congenital and acquired factor VIII or IX inhibitors.. J Thromb Haemost 2004; 2: 899-909.
• 2 Boffard KD, Riou B, Warren B. et al. Recombinant factor VIIa as adjunctive therapy for bleeding control in severely injured trauma patients: two parallel randomized, placebo-controlled, double-blind clinical trials.. J Trauma 2005; 59: 8-15.
• 3 Grounds RM, Bolan C. Clinical experiences and current evidence for therapeutic recombinant factor VIIa treatment in nontrauma settings.. Crit Care 2005; 9 (Suppl. 05) S29-S36.
• 4 Diprose P, Herbertson MJ, O’Shaughnessy D. et al. Activated recombinant factor VII after cardiopulmonary bypass reduces allogeneic transfusion in complex non-coronary cardiac surgery: randomized double-blind placebo-controlled pilot study.. Br J Anaesth 2005; 95: 596-602.
• 5 Planinsic RM, van der Meer J, Testa G. et al. Safety and efficacy of a single bolus administration of recombinant factor VIIa in liver transplantation due to chronic liver disease.. Liver Transpl 2005; 11: 895-900.
• 6 Lodge JP, Jonas S, Jones RM. et al; rFVIIa OLT Study Group. Efficacy and safety of repeated perioperative doses of recombinant factor VIIa in liver transplantation.. Liver Transpl 2005; 11: 973-979.
• 7 Friederich PW, Henny CP, Messelink EJ. et al. Effect of recombinant activated factor VII on perioperative blood loss in patients undergoing retropubic prostatectomy: a double-blind placebo-controlled randomised trial.. Lancet 2003; 361: 201-205.
• 8 Ahonen J, Jokela R. Recombinant factor VIIa for life-threatening post-partum haemorrhage.. Br J Anaesth 2005; 94: 592-595.
• 9 O’Connell NM, Perry DJ, Hodgson AJ. et al. Recombinant FVIIa in the management of uncontrolled hemorrhage.. Transfusion 2003; 43: 1711-1716.
• 10 Monroe DM, Hoffman M, Oliver JA. et al. Platelet activity of high-dose factor VIIa is independent of tissue factor.. Br J Haematol 1997; 99: 542-547.
• 11 Persson E, Kjalke M, Olsen OH. Rational design of coagulation factor VIIa variants with substantially increased intrinsic activity.. Proc Natl Acad Sci USA 2001; 98: 13583-13588.
• 12 Tranholm M, Kristensen K, Kristensen AT. et al. Improved hemostasis with superactive analogs of factor VIIa in a mouse model of hemophilia A.. Blood 2003; 102: 3615-3620.
• 13 Ghosh S, Ezban M, Persson E. et al. Activity and regulation of factor VIIa analogs with increased potency at the endothelial cell surface.. J Thromb Haemost 2007; 5: 336-346.
• 14 Allen GA, Persson E, Campbell RA. et al. A variant of recombinant factor VIIa with enhanced procoagulant and antifibrinolytic activities in an in vitro model of hemophilia.. Arterioscler Thromb Vasc Biol 2007; 27: 683-689.
• 15 Lisman T, de Groot PG, Lambert T. et al. Enhanced in vitro procoagulant and antifibrinolytic potential of superactive variants of recombinant factor VIIa in severe hemophilia A.. J Thromb Haemost 2003; 1: 2175-2178.
• 16 Nicolaisen EM. Long-term follow-up with regard to potential immunogenicity: Clinical experience with NovoSeven® (recombinant factor VIIa).. Haemostasis 1996; 26 (Suppl. 01) 98-101.
• 17 Pittman DD, Alderman EM, Tomkinson KN. et al. Biochemical, immunological, and in vivo functional characterization of B-domain-deleted factor VIII.. Blood 1993; 81: 2925-2935.
• 18 Nicolaisen EM, Kristensen H, Kristensen A. et al. Evaluation of potential antigenicity of active-site-inhibited recombinant human FVIIa (FFR-rFVIIa) in an immune-tolerant rat model.. Thromb Haemost 2002; 87: 836-839.
• 19 ICH Harmonised Tripartite Guideline: Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals S6 (Ref. CPMP/ICH/302/95). ICH 1997. http://www.ich.org/LOB/media/MEDIA503.pdf
• 20 Committee for Medicinal Products for Human Use.. Concept Paper on Guideline on Immunogenicity Assessment of Biotechnology-Derived Therapeutic Proteins (Ref. EMEA/CHMP/BMWP/246511/2005). EMEA 2007. http://www.emea.europa.eu/pdfs/human/biosimilar/24651105en.pdf
• 21 Persson E, Bak H, Olsen OH. Substitution of valine for leucine 305 in factor VIIa increases the intrinsic amidolytic activity.. J Biol Chem 2001; 276: 29195-29199.
• 22 Lev EI, Marmur JD, Zdravkovic M. et al. Antithrombotic effect of tissue factor inhibition by inactivated factor VIIa: an ex vivo human study.. Anterioscler Thromb Vasc Biol 2002; 22: 1036-1041.