title: 2-(2-Br-phenyl)-8-methoxy-benzoxazinone (HPW-RX2), a direct thrombin inhibitor with a suppressive effect on thromboxane formation in platelets.
abstract: 2-(2-Br-phenyl)-8-methoxy-benzoxazinone (HPW-RX2), a newly synthetic benzoxazinone derivative, has previously been shown to inhibit rabbit platelet aggregation caused by thrombin and arachidonic acid. In the present study, the mechanism for the antiplatelet effect of HPW-RX2 was further investigated. In human platelets, HPW-RX2 concentration-dependently inhibited platelet aggregation, ATP release, P-selectin expression, and intracellular calcium mobilization caused by thrombin. In contrast, HPW-RX2 had no significant effect on either SFLLRN- or GYPGKF-induced platelet aggregation, indicating that HPW-RX2 did not interfere with platelet thrombin receptors. Moreover, HPW-RX2 inhibited the amidolytic activity of thrombin and prolonged the fibrinogen clotting time. These results suggest that the inhibitory effect of HPW-RX2 on thrombin-induced platelet aggregation is via direct inhibition of thrombin proteolytic activity. Besides the inhibition on thrombin, HPW-RX2 also prevented platelet aggregation, ATP release, and increase in [Ca2+]i caused by arachidonic acid and low concentration collagen. In a parallel manner, both arachidonic acid-induced thromboxane B2 and prostaglandin D2 formations were decreased in platelets treated with HPW-RX2. This indicates that HPW-RX2 is able to inhibit the arachidonic acid cascade at the cyclooxygenase level. This is the first report of a benzoxazinone derivative possessing both thrombin and cyclooxygenase inhibitory properties. The dual effect of HPW-RX2 might provide extra therapeutic benefits for treatment of arterial thrombosis.
authors.affiliation: Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, Taiwan. ccwu@kmu.edu.tw
authors.forename: Chin-Chung
authors.initials: CC
authors.lastname: Wu
authors.forename: Tsai-Wei
authors.initials: TW
authors.lastname: Wang
authors.forename: Wei-Ya
authors.initials: WY
authors.lastname: Wang
authors.forename: Pei-Wen
authors.initials: PW
authors.lastname: Hsieh
authors.forename: Yang-Chang
authors.initials: YC
authors.lastname: Wu
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism / Q000494:pharmacology; D001241:Aspirin / Q000494:pharmacology; D048588:Benzoxazines / Q000737:chemistry / Q000494:pharmacology*; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D019007:P-Selectin / Q000096:biosynthesis; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000494:pharmacology; D015230:Prostaglandin D2 / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D018179:Receptors, Thrombin / Q000378:metabolism; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000494:pharmacology; D013928:Thromboxane A2 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D013929:Thromboxane B2 / Q000037:antagonists & inhibitors / Q000096:biosynthesis
chemical_list: C507651:2-(2-Br-phenyl)-8-methoxy-benzoxazinone; D048588:Benzoxazines; D019007:P-Selectin; D010975:Platelet Aggregation Inhibitors; D018179:Receptors, Thrombin; D016718:Arachidonic Acid; D013929:Thromboxane B2; D013928:Thromboxane A2; D000255:Adenosine Triphosphate; D003094:Collagen; D013917:Thrombin; D001241:Aspirin; D015230:Prostaglandin D2; D002118:Calcium