Hamdan Medical Journal (previously the Journal of Medical Sciences)

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Modulation of Serine Proteases-Mediated Platelet Activation

Sarfraz Ahmad
Published in : Journal of Medical Sciences ; Vol 3, No 1 (2010)
DOI : 10.2174/1996327001003010044


Novel direct thrombin inhibitors (DTIs), such as bivalirudin, are replacing heparin in several clinical scenarios. In particular, DTIs are indicated for the treatment and thromboprophylaxis of patients with heparin-induced thrombocytopenia (HIT). In interventional cardiology, DTIs have several advantages over heparin, and offer a clinical benefit equivalent to that of a combination of heparin and antiplatelet agents. We hypothesize that this benefit results from the ability of DTIs to inhibit platelet activation by activated serine proteases. This study represents the development of a modified 14C-serotonin release assay (SRA) to investigate the relative inhibitory effects of three DTIs (argatroban, bivalirudin and hirudin) on thrombin- and factor Xa-mediated 14C-serotonin release (SR) in plasma-free systems. Washed platelets were isolated from blood of healthy volunteers. The 14CSRA test was similar to that used to detect heparin-PF4 antibody-mediated platelet activation, except that it was used to evaluate the ability of DTIs to modulate protease-induced SR responses. The inhibitory effects of DTIs were determined at protease concentrations that induced ≥50% SR. Serine proteases induced SR from platelets in a concentration-dependent manner. Human thrombin was found to be more potent than bovine thrombin (2-3 times for 50-80% SR). Bovine factor Xa (≥0.2 nKat/ml) produced a comparable (50- 80%) SR response. All three DTIs effectively blocked serine protease-mediated platelet activation in a concentrationdependent manner. The optimum inhibitory concentrations of bivalirudin on SR was ~100 nM for human thrombin and bovine factor Xa and almost double for bovine thrombin; well below plasma concentrations necessary for effective anticoagulation for percutaneous coronary interventions. Wide variations in the inhibitory effects of each DTI on thrombin- and factor Xa-mediated platelet activation were noted, which was partly dependent on the donor platelets and stability of the proteases/inhibitors. It is concluded that DTIs can directly inhibit serine proteases- mediated platelet activation responses.

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