A Human Antibody that Binds to the γ-carboxyglutamic Acid Domain of Factor IX Is a Potent Antithrombotic In Vivo

Canio J. Refino; Jacques Himber; Louis Burcklen; Paul Moran; Mark Peek; Shelley Suggett; Brigitte Devaux; Daniel Kirchhofer

doi:10.1055/s-0037-1614351

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1999; 82(03): 1188-1195
DOI: 10.1055/s-0037-1614351

Letters to the Editor

Schattauer GmbH

A Human Antibody that Binds to the γ-carboxyglutamic Acid Domain of Factor IX Is a Potent Antithrombotic In Vivo

Authors

Canio J. Refino

¹From the Genentech Inc., Cardiovascular Research and Antibody Technologies Departments, South San Francisco, CA, USA
Jacques Himber

²F. Hoffmann-La Roche, Preclinical Research, Basel, Switzerland
Louis Burcklen

²F. Hoffmann-La Roche, Preclinical Research, Basel, Switzerland
Paul Moran

¹From the Genentech Inc., Cardiovascular Research and Antibody Technologies Departments, South San Francisco, CA, USA
Mark Peek

¹From the Genentech Inc., Cardiovascular Research and Antibody Technologies Departments, South San Francisco, CA, USA
Shelley Suggett

¹From the Genentech Inc., Cardiovascular Research and Antibody Technologies Departments, South San Francisco, CA, USA
Brigitte Devaux

¹From the Genentech Inc., Cardiovascular Research and Antibody Technologies Departments, South San Francisco, CA, USA
Daniel Kirchhofer

¹From the Genentech Inc., Cardiovascular Research and Antibody Technologies Departments, South San Francisco, CA, USA

Abstract

Summary

10C12, a human antibody F(ab’)₂, which specifically binds to the Gla domain of factor IX, interfered with all known coagulation processes that involve factor IX/IXa. These include the function of the intrinsic Xase complex and the activation of zymogen factor IX by factor XIa and by the tissue factor:factor VIIa complex. Furthermore, 10C12 potently inhibited activated partial thromboplastin clotting times (APTT) in plasma of guinea pig and rat, thus enabling in-vivo evaluation. In guinea pigs, a bolus administration of 10C12 (10 μg/kg) prevented cyclic flow variations in damaged carotid arteries without affecting coagulation or bleeding parameters. At a 100-fold higher dose, 10C12 had no effect on normal hemostasis as assessed by the cuticle bleeding time. At this dose, 10C12 was also efficacious in a rat arterial thrombosis model, substantially reducing clot weight and duration of vessel occlusion while prolonging ex-vivo APTT only 1.2-fold. The dose of heparin required to produce comparable antithrombotic effects prolonged the APTT by 12-fold and increased the tail bleeding time (TBT) by 8-fold. In contrast, 10C12 had no effect on TBT. However, rat tails showed a tendency for rebleeding which 10C12 exacerbated. In conclusion, the antithrombotic potency of the 10C12 antibody in two species provides evidence for an important role of F.IX, and its Gla domain in particular, during thrombogenesis under arterial flow conditions. The relative safety at effective doses of this fully human antibody suggests that it may have therapeutic value for treatment of thrombotic disorders.

Abbreviations

Gla: γ-carboxyglutamic acid; CDRs: complementarity determining regions; CVFs: cyclic flow variations; TF:F.VIIa: tissue factor:factor VIIa; APTT: activated partial thromboplastin time; PT: prothrombin time; ACT: whole blood activated clotting time; CBT: cuticle bleeding time; TBT: tail bleeding time

Full Text

References

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