Thromb Haemost 2000; 84(05): 912-917
DOI: 10.1055/s-0037-1614136
Review Article
Schattauer GmbH

S-Nitrosoderivative of a Recombinant Fragment of von Willebrand Factor (S-Nitroso-AR545C) Inhibits Thrombus Formation in Guinea Pig Carotid Artery Thrombosis Model

Osnat Gurevitz
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
Michael Eldar
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
Ehud Skutelsky
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
Ilia Tamarin
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
Boris Shenkman
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
Regina Eskaraev
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
David Castel
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
Joseph Loscalzo
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
,
Aida Inbal
1   From the Neufeld Cardiac Research and Thrombosis and Hemostasis Institutes, Sheba Medical Center, Tel Hashomer and Pathology Department, the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel; and the Whitaker Cardiovascular Institute and Evans Department of Medicine, Boston University School of Medicine, Boston, MA, USA
› Author Affiliations

This study was supported in part by Shlezak Grant from Tel Aviv University, Israel and NitroMed company, Boston MA, USA. We are indebted to Mrs. Elaine Finkelstein for excellent secretarial assistance.
Further Information

Publication History

Received 22 February 2000

Accepted after resubmission 22 May 2000

Publication Date:
13 December 2017 (online)

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Summary

Antiplatelet drugs are the mainstays of therapy for acute and chronic cardiovascular diseases. S-nitroso-AR545C – an S-nitrosoderivative of a recombinant von Willebrand factor fragment AR545C spanning Ala 444 to Asp 730 and containing an Arg 545 Cys mutation, was previously found to inhibit ristocetin-and ADP-induced platelet aggregation and the interaction of platelets with extracellular matrix (ECM). In the current study we tested the antithrombotic properties of S-nitroso-AR545C on guinea pig platelets and in a platelet-rich thrombosis model in the guinea pig. Preincubation of guinea pig platelets with 0.1 µM of S-nitroso-AR545C decreased ristocetin-induced agglutination by 40% (p = 0.009) and completely abolished ADP-induced aggregation (p <0.0001). At concentration of 1.0 µM, S-nitroso-AR545C completely inhibited platelet adhesion (represented by surface coverage – SC) and decreased aggregate formation (represented by average aggregate size – AS) by more than 50%. Treatment of guinea pigs with 1.0 mg/kg S-nitroso-AR545C resulted in a significantly delayed time to arterial occlusion (31.7 ± 6.0 min vs. 13.9 ± 3.2 min, p <0.02). Similarly, total patency time was longer in the group injected with S-nitroso-AR545C compared to the control group. However, the difference was not statistically significant (33.8 ± 6.3 min vs. 20.2 ± 3.3 min, p = 0.07). No change in platelet count, hematocrit and bleeding time was observed 60 min after injection compared to baseline. In contrast, a significant decrease in SC (p <0.0001) and AS (p <0.01) were observed 60 min after the injection of S-nitroso-AR545C, whereas no change in these parameters was observed in the control group. These observations indicate that S-nitroso-AR545C exhibits significant antiadhesive and antiaggregating effects in-vitro and inhibits clot formation in-vivo suggesting that this compound may have potential therapeutic advantages.