Comparison of Recombinant Human Thrombin and Plasma-Derived Human α-Thrombin

ABSTRACT

Bleeding can be a serious complication of surgery, and topical thrombin is widely used as an adjunct to hemostasis in diverse surgical settings. The potent hemostatic properties of thrombin derive from its ability to activate platelets directly to aggregate and adhere to damaged vessels and to catalyze the formation simultaneously of a fibrin matrix. Application of exogenous thrombin bypasses the physiological process of generating a thrombin burst by directly initiating the terminal reactions of blood clot formation. Currently, thrombin used to control surgical bleeding is primarily from bovine plasma, with a small percentage from human plasma. Human thrombin isolated from pooled plasma carries the risk of transmitting plasma-borne pathogens or prion diseases. The bovine preparations have been associated with protein and preparative contaminants that pose potential risks of developing cross-reacting antibodies. There is a need for a pure therapeutic preparation of human thrombin. Recombinant human thrombin (rhThrombin) has been efficiently produced from a prethrombin-1 precursor obtained from Chinese hamster ovary cell culture. This rhThrombin is substantially free of process-derived contaminants and has been characterized extensively in terms of composition, primary, secondary, and tertiary structure, enzymatic activity; and in vivo pharmacology. In vivo studies of topically applied rhThrombin have shown it is effective in achieving hemostasis in a rabbit liver excisional wound model. Clinical studies are ongoing to evaluate the safety and efficacy of rhThrombin as an adjunct to hemostasis in patients undergoing surgery.

rhThrombin - prethrombin - prothrombin

REFERENCES

• 1 Hedner U. Mechanism of action of factor VIIa in the treatment of coagulopathies.  Semin Thromb Hemost. 2006;  32(suppl 1) 77-85
• 2 Roberts H, Hoffman M, Monroe D. A cell-based model of thrombin generation.  Semin Thromb Hemost. 2006;  32(suppl 1) 32-38
• 3 Schoenecker J G, Hauck R K, Mercer M C, Parker W, Lawson J H. Exposure to topical bovine thrombin during surgery elicits a response against the xenogeneic carbohydrate galactose alpha1-3galactose.  J Clin Immunol. 2000;  20 434-444
• 4 Bishop P, Lawson J. Recombinant biologics for treatment of bleeding disorders.  Nat Rev Drug Discov. 2004;  3 684-694
• 5 Ortel T L, Mercer M C, Thames E H, Moore K D, Lawson J H. Immunologic impact and clinical outcomes after surgical exposure to bovine thrombin.  Ann Surg. 2001;  233 88-96
• 6 Winterbottom N, Kuo J M, Nguyen K et al.. Antigenic responses to bovine thrombin exposure during surgery: a prospective study of 309 patients.  J Appl Res. 2002;  2 1-11
• 7 Lawson J. The clinical use and immunologic impact of thrombin in surgery.  Semin Thromb Hemost. 2006;  32(suppl 1) 98-110
• 8 Franchini M, Zaffanello M, Veneri D. Recombinant factor VIIa. An update on its clinical use.  Thromb Haemost. 2005;  93 1027-1035
• 9 Pipe S W. The promise and challenges of bioengineered recombinant clotting factors.  J Thromb Haemost. 2005;  3 1692-1701
• 10 Blostein M D, Furie B C, Rajotte I, Furie B. The Gla domain of factor IXa binds to factor VIIIa in the tenase complex.  J Biol Chem. 2003;  278 31297-31302
• 11 Sugiura T, Maruyama H B. Factors influencing expression and post-translational modification of recombinant protein C.  J Biotechnol. 1992;  22 353-360
• 12 Bendig M M. The production of foreign proteins in mammalian cells.  Genet Eng. 1988;  7 91-127
• 13 Jenny N, Mann K. Thrombin. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN Hemostasis and Thrombosis: Basic Principles and Clinical Practice. Philadelphia, PA; Lippincott Williams & Wilkins 2001
• 14 Liu L W, Vu T K, Esmon C T, Coughlin S R. The region of the thrombin receptor resembling hirudin binds to thrombin and alters enzyme specificity.  J Biol Chem. 1991;  266 16977-16980
• 15 Möller T P, Weinstein J R, Hanisch U-K. Activation of microglial cells by thrombin: past, present, and future.  Semin Thromb Hemost. 2006;  32(suppl 1) 69-76
• 16 Whitton C, Sands D, Lee T, Chang A, Longstaff C. A reunification of the US (“NIH”) and International Unit into a single standard for thrombin.  Thromb Haemost. 2005;  93 261-266
• 17 Sonder S A, Fenton J W. Thrombin specificity with tripeptide chromogenic substrates: comparison of human and bovine thrombins with and without fibrinogen clotting activities.  Clin Chem. 1986;  32 934-937
• 18 Hemker H C, Beguin S. Thrombin generation in plasma: its assessment via the endogenous thrombin potential.  Thromb Haemost. 1995;  74 134-138