Pre-Clinical Characteristics of a Recombinant Fusion Protein Linking Activated Coagulation Factor VII with Albumin (rVIIa-FP)
Sabine ZollnerDaniel SchuermannElmar RaquetJochen Mueller‐CohrsThomas WeimerIngo PragstGerhard DickneiteStefan Schulte
0
Citation
0
Reference
10
Related Paper
Keywords:
Recombinant Factor VIIa
Factor IX
Pharmacodynamics
Factor VII
Factor VII
Factor IX
Factor VIIa
Clotting factor
Clotting time
Factor IXa
Factor V
Cite
Citations (72)
-The development of factor VIII and factor IX concentrates markedly improved the management of hemophilia A and B and made home therapy possible. However, treatment of hemophilic patients with acquired inhibitors remained difficult. The preparation of prothrombin complex concentrates, nonactivated or activated, addressed this difficult clinical problem, but their use was associated with serious side effects. Factor VIIa was found to be a safer treatment modality. Factor VIIa per se is inactive and needs tissue factor (TF) to become biologically active. TF serves as the receptor for factor VIIa. TF is expressed from vessel walls only upon injury. Treatment of inhibitor patients with plasma-derived factor VII was found to be successful. This led to the development of recombinant factor VIIa, which was also found to be successful in managing hemophilia patients with inhibitors. Up to this point, large numbers of patients have been treated and few serious side effects have been noted. Because of its unique effects on the hemostasis system, recombinant factor VIIa will be useful for other indications as well, including patients with congenital factor VII deficiency, patients with bleeding and liver function impairment, patients with quantitative and qualitative platelet defects, and individuals who have sustained multiple trauma. The development of recombinant factor VII is reviewed in detail.
Recombinant Factor VIIa
Factor VII
Factor IX
Factor VIIa
Prothrombin complex concentrate
Cite
Citations (77)
Factor VII
Factor IX
Factor VIIa
Clotting factor
Clotting time
Factor V
Cite
Citations (4)
An abnormal blood coagulation factor IX has been isolated from the blood of a hemophilia B patient with a variant of the disease (hemophilia Bm) characterized by a normal concentration of factor IX antigen, negligible factor IX coagulant activity, and a prolonged prothrombin time with bovine tissue factor. The isolated protein (factor IXBm) had the same apparent molecular weight as normal factor IX (55,000) and the same mobility on two dimensional immunoelectrophoresis as normal factor IX. Factor IXBm underwent limited proteolysis induced by activated factor XI, in the presence of Ca2+ ions, or induced by the reaction product of tissue factor, factor VII and Ca2+ ions. A timecourse study showed that activated factor XI cleaved factor IXBm and factor IX at similar rates. However, in contrast to normal factor IX, the limited protelysis of factor IXBm did not generate procoagulant activity. In kinetic experiments purified factor IXBm behaved like a competitive inhibitor (Ki of 0.017 muM) of the activation of factor X by bovine tissue factor and factor VII. Normal factor IX was also found to inhibit the reaction but required a four-fold higher concentration to activate the same inhibitory effects as factor IXBm.
Factor IX
Factor VII
Factor IXa
Thromboplastin
Factor V
Cite
Citations (29)
The vitamin K-dependent plasma proteins demonstrate remarkable similarities in their structures: all have multiple domains in common and extensive homology is observed within many of these domains. In order to investigate the structure-function relationship of these proteins, we have interchanged domains of one protein (factor IX) with that of another (factor VII) and have compared the expression of these fusion proteins with recombinant and native factors IX and VII. Oligonucleotide-directed mutagenesis was used to generate four fusion proteins: factor IX/VII-1, which contains the factor IX leader and gla domain fused to the growth factor and serine protease of factor VII; factor VII/IX-1, a reciprocal fusion protein of factor IX/VII-1; factor IX/VII-2, which contains the factor IX leader adjoined to the mature factor VII protein sequence; and factor VII/IX-2, the reciprocal fusion protein of factor IX/VII-2. The cDNAs encoding all four proteins were cloned into mammalian expression vectors, and to date three of these (factors IX/VII-1, 2 and VII/IX-1) have been transfected into baby hamster kidney (BHK) cells or 293 cells and characterized. Factors IX/VII-1 and VII/IX-1 were both secreted at levels comparable to recombinant factors IX and VII. The factor IX/VII-1 was identical in molecular weight to native or recombinant factor VII (i.e., 53 K). Factor VII/IX-1 was expressed as two proteins with molecular weights around 68 kd, as observed with recombinant factor IX. The factor IX/VII-1 protein has been purified to homogeneity and has been found to possess factor VII biological activity, but at a specific activity approximately 20% that of plasma factor VII. Thus, the gla domain of one clotting factor is capable of directing the activation of another and of generating biologically active protein. In contrast, no activity was observed with the factor IX/VII-2 fusion protein, indicating that there are limits to the interchanges which can generate functional blood clotting factors.
Factor IX
Factor VII
Cite
Citations (0)
Factor VII deficiency is a rare autosomal bleeding disorder with a highly variable hemorrhagic predisposition. Severe bleeding, including hemarthroses, may be encountered when plasma factor VII levels are below 1%. Patients have prolonged prothrombin times, and the final diagnosis is established by quantitative factor VII assays. Some patients have true deficiencies, that is, very low factor VII activity and low factor VII antigen (cross-reacting material) levels (CRM-); others have normal antigen levels but low activity (CRM+). Still others have reduced antigen levels (CRMR). There is a rather poor correlation between clinical symptoms and factor VII activity levels in plasma. Treatment of these patients consists of fresh frozen plasma, prothrombin complex concentrates, or factor VII concentrates. Recombinant activated factor VII (rFVIIa) is a very useful alternative, and several patients have been treated successfully. Because of the short half-life of factor VIIa, repeated doses have to be administered, and continuous infusion may be even better. Antibodies to factor VII have been reported but seem to be rather rare. From the available data it appears that rFVIIa is a safe and effective treatment modality for patients with congenital factor VII deficiency.
Factor VII
Recombinant Factor VIIa
Fresh frozen plasma
Prothrombin complex concentrate
Factor VIIa
PROTHROMBIN COMPLEX
Prothrombin time
Cite
Citations (74)
Factor IX
Factor VII
Thrombogenicity
Thromboplastin
Sephadex
Clotting factor
Cite
Citations (2)
Purpose of review To examine the development of recombinant FVIIa (rFVIIa); a new concept of inducing hemostasis. It was developed for use in hemophilia patients with inhibitors against FVIII or FIX with the vision to provide these patients with a therapeutical option to be used instead of FVIII or FIX. For the first time it was shown that pharmacological doses of FVIIa induced hemostasis. Recent findings Hemostasis was achieved by rFVIIa in major surgery (repeated doses) as well as in a home-treatment setting (one single injection) in severe hemophilia patients with inhibitors. A recent study indicates that rFVIIa may be useful as prophylaxis. In heavily bleeding nonhemophilia patients rFVIIa was shown to induce hemostasis. Pharmacological doses of rFVIIa enhance thrombin generation on activated platelets resulting in the formation of tight hemostatic fibrin plugs resistant against premature proteolysis. High doses of rFVIIa seem to be safe probably due to its localized effect. Summary Pharmacological doses of rFVIIa induce hemostasis in severe hemophilia and in nonhemophilia patients with profuse, heavy bleeding. rFVIIa enhances thrombin generation on activated platelets, thereby initiating the formation of strong, tight fibrin hemostatic plugs resistant to premature lysis. It also seems to be safe in high doses.
Recombinant Factor VIIa
Thrombin Generation
Hematology
Cite
Citations (50)
Six brands of Factor IX concentrates were evaluated for their Factor VII content and for the presence of activated Factor VII through use of a coupled amidolytic assay, insensitive to activated Factor VII, and a clotting assay, sensitive to activated Factor VII. The Factor VII content of the concentrates studied (except for one concentrate purposely produced to exclude Factor VII) varied between 33 to 621 U per vial. All concentrates contained activated Factor VII, as indicated by ratios of Factor VII clotting activity to Factor VII amidolytic activity of from 1.6 to 21.5. Higher ratios were found in two brands of activated concentrates than in non-activated concentrates. In 10 patients infused with Factor IX concentrates, plasma Factor VII activity rose strikingly in the clotting assay but not in the amidolytic assay. Thus, the elevated Factor VII levels by the clotting assay after infusion of Factor IX concentrates stem from circulating activated Factor VII. A mean intravascular half-disappearance time of 144 min was found for activated Factor VII. Its persistence in the circulation makes it important to evaluate the possible role of activated Factor VII in the thrombogenicity of Factor IX concentrates and in their reported effectiveness in treating bleeding in Hemophilia A patients with inhibitors.
Factor IX
Factor VII
Thrombogenicity
Clotting factor
Clotting time
Cite
Citations (2)
Factor IX
Thrombogenicity
Factor VII
Thromboplastin
Sephadex
Clotting factor
Cite
Citations (67)