Comparison of High Purity Factor IX Concentrates and a Prothrombin Complex Concentrate in a Canine Model of Thrombogenicity

Summary

A non-stasis canine model of thrombogenicity has been used to evaluate batches of high purity factor IX concentrates from 4 manufacturers and a conventional prothrombin complex concentrate (PCC). Platelets, activated partial thromboplastin time (APTT), fibrinogen, fibrin(ogen) degradation products and fibrinopeptide A (FPA) were monitored before and after infusion of concentrate. Changes in FPA were found to be the most sensitive and reproducible indicator of thrombogenicity after infusion of batches of the PCC at doses of between 60 and 180 IU/kg, with a dose related delayed increase in FPA occurring. Total FPA generated after 100-120 IU/kg of 3 batches of PCC over the 3 h time course was 9-12 times that generated after albumin infusion. In contrast the amounts of FPA generated after 200 IU/kg of the 4 high purity factor IX products were in all cases similar to albumin infusion. It was noted that some batches of high purity concentrates had short NAPTTs indicating that current in vitro tests for potential thrombogenicity may be misleading in predicting the effects of these concentrates in vivo.

• References

• 1 Aronson DL. Factor IX complex. Semin Thromb Hemostas 1979; 1: 28-43
• 2 Middleton SM, Bennet IH, Smith JK. A therapeutic concentrate of coagulation factors II, IX and X from citrated factor VIII depleted plasma. Vox Sang 1973; 24: 441-56
• 3 Dike GWR, Bidwell E, Rizza CR. The preparation and clinical use of a new concentrate containing factor IX, prothrombin and factor X, and of a separate concentrate containing factor VII. Br J Haematol 1972; 22: 469-90
• 4 Heystek J, Brummelhuis HGJ, Krinjen HW. Contribution to the optimal use of human blood. II. The large-scale preparation of prothrombin complex. A comparison between two methods using the anion exchangers DEAE-cellulose DE-52 and DEAE-Sephadex A-50. Vox Sang 1973; 25: 113-23
• 5 Hoag MS. Clinical experience with a new clotting factor concentrate. Thromb Diath Haemorrh 1970; Suppl 39 417-20
• 6 Seligsohn U, Kasper CK, Osterud B, Rapaport SI. Activated factor VII: Presence in factor IX concentrates and persistence in the circulation after infusion. Blood 1979; 53: 828-37
• 7 Mannucci PM, Vigano S. Protein C concentrates for clinical use. Lancet 1983; I: 875
• 8 Blatt PM, Kingdon HS, McLean G, Roberts HR. Thrombogenic material in prothrombin complex concentrates. Ann Intern Med 1974; 81: 766-70
• 9 Marassi A, Manzullo V, di Carlo V, Mannucci PM. Thromboembolism following prothrombin complex concentrates and major surgery in liver disease. Thromb Haemostas 1978; 39: 787-8
• 10 Kasper CK. Thromboembolic complications. Thromb Diath Haemorrh 1975; 33: 640-4
• 11 Magner A, Aronson DL. Toxicity of factor IX concentrates in mice. Dev Biol Stand 1979; 44: 185-8
• 12 Hultin MB. Activated clotting factors in factor IX concentrates. Blood 1979; 54: 1028-38
• 13 Giles AR, Nesheim ME, Hoogendoorn H, Tracy PB, Mann KG. The coagulant-active phospholipid content is a major determinant of in vivo thrombogenicity of prothrombin complex (factor IX) concentrates in rabbits. Blood 1982; 59: 401-7
• 14 Prowse CV, Chirnside A, Elton R. In vitro thrombogenicity tests of factor IX concentrates. I. A survey of available assays. Thromb Haemostas 1979; 42: 1355-67
• 15 Kingdon HS, Lundblad RL, Veltkamp JJ, Aronson DL. Potentially thrombogenic materials in factor IX concentrates. Thromb Diath Haemorrh 1975; 33: 617-31
• 16 Cash JD, Dalton RG, Middleton S, Smith JK. Studies on the thrombogenicity of Scottish factor IX concentrates in dogs. Thromb Diath Haemorrh 1975; 33: 632-9
• 17 Hedner U, Nilsson IM, Bergentz S-E. Various prothrombin complex concentrates and their effect upon coagulation and fibrinolysis in vivo. Thromb Haemostas 1976; 35: 386-95
• 18 Burnouf T, Michalski C, Goudemand M, Huart JJ. Properties of a highly purified human plasma factor IX:c therapeutic concentrate prepared by conventional chromatography. Vox Sang 1989; 57: 225-32
• 19 Menache D, Behre HE, Orthner CL, Nunez H, Anderson HD, Triantaphyllopoulos DC, Kosow DP. Coagulation factor IX concentrate: method of preparation and assessment of potential in vivo thrombogenicity in animal models. Blood 1984; 64: 1220-7
• 20 Mannucci PM, Bauer KA, Gringeri A, Barzegar S, Bottasso B, Simoni L, Rosenberg RD. Thrombin generation is not increased in the blood of hemophilia B patients after the infusion of a purified factor IX concentrate. Blood 1990; 76: 2540-5
• 21 Kim HC, McMillan CW, White GC, Bergman GE, Saidi P. Clinical experience of a new monoclonal antibody purified factor IX: half-life, recovery, and safety in patients with haemophilia B. Sem Haematol 1990; 27 (Suppl 2) 30-5
• 22 Littlewood JD, Dawes J, Smith JK, Feldman PA, Haddon ME, McQuillan TA, Foster PR, Ferguson JM, Prowse CV. Studies on the effect of heat treatment on the thrombogenicity of factor IX concentrates in dogs. Br J Haematol 1987; 65: 463-8
• 23 MacGregor IR, Ferguson JM, Dawes J, McLaughlin L, Prowse CV. Thrombogenicity of a factor IX concentrate quantitated in a canine model. Blood Coag Fibrinol 1990; 1: 23-30
• 24 Sas G, Owens RE, Smith JK, Middleton S, Cash JD. In vitro spontaneous thrombin generation in human factor IX concentrates. Br J Haematol 1975; 31: 25-35
• 25 Nesheim ME, Taswell JB, Mann KG. The contribution of bovine factor V and factor Va to the activity of prothrombinase. J Biol Chem 1979; 254: 10952-62
• 26 Gornal AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem 1949; 177: 751-66
• 27 Wilner GD. Measurement of fibrinopeptide A in canine blood: an interim report. Thromb Res 1979; 15: 601-10
• 28 Mannucci PM, Bauer KA, Gringeri A, Barzegar S, Santagostino E, Tardati FC, Rosbenberg RD. No activation of the coagulation cascade after large doses of a high-purity Factor IX concentrate (submitted for publication).
• 29 Giles AR, Nesheim ME, Hoogendoorn H, Tracy PB, Mann KG. Stroma free human platelet lysates potentiate the in vivo thrombogenicity of factor Xa by the provision of coagulant-active phospholipid. Br J Haematol 1982; 51: 457-68
• 30 Prowse C, Pepper DS. In vitro tests of the potential thrombogenicity of factor IX concentrates: inhibition and characterisation studies of NAPTT, TGt50 and PF3 moieties. Thromb Res 1980; 20: 491-8