Wu C.-M.Lin K.-W.Teng C.-H.Huang A.-M.Chen Y.-C.Yen M.-H.Wu W.-B.YEONG-SHIAU PULin C.-N.2021-02-022021-02-0220140918-6158https://www.scopus.com/inward/record.uri?eid=2-s2.0-84904673676&doi=10.1248%2fbpb.b14-00099&partnerID=40&md5=1b49a356525493e783f55b65105d35a8https://scholars.lib.ntu.edu.tw/handle/123456789/544382In an effort to develop potent cyclooxygenase-1 (COX-1) inhibitors used as anticancer agent, a series of 2′,5′-dimethoxychalcones was screened to evaluate their antiplatelet effect on human washed platelets suspension. Compound 2 exhibited potent inhibition of human washed platelet aggregation induced by collagen, significantly inhibited collagen- and arachidonic acid-induced thromboxane B2 release, and revealed inhibitory effect on COX-1 activity. Molecular docking studies showed that 1, 2, and 4 were bound in the active site of COX-1. These indicated that the antiplatelet effect of these compounds were mainly mediated through the suppression of COX-1 activity and reduced the thromboxane formation. To investigate the mechanistic action of COX-1 inhibitor enhanced the cytotoxic effect against human bladder cancer cells, NTUB1, we assessed the cytotoxic effect of 2 against NTUB1. Treatment of NTUB1 cells with various concentrations of 2 led to a concentration-dependent increase of cell death and decrease of reactive oxygen species levels. The flow-cytometric analysis showed that 2 induced a G1 phase cell cycle arrest but did not accompany an appreciable sub-G1 phase in NTUB1 cells. In addition, compound 2 increased p21 and p27 expressions and did not inhibit the expression of COX-1 in NTUB1 cells. Our results suggested that 2 enhanced cell growth inhibition or antiproliferative activity in NTUB1 cells through G1 arrest by COX-1 independent mechanism. ? 2014 The Pharmaceutical Society of Japan.Antiplatelet; Bladder cancer; Chalcone; Cyclooxygenase-1 (COX-1); Cytotoxicity; G1 cell cycle arrest[SDGs]SDG32',5' dimethoxychalcone derivative; chalcone derivative; collagen; cyclooxygenase 1; protein p21; protein p27; reactive oxygen metabolite; thromboxane B2; unclassified drug; antineoplastic agent; antithrombocytic agent; chalcone; cyclooxygenase 1; reactive oxygen metabolite; article; bladder cancer; cancer cell; cancer cell line; cancer inhibition; cell cycle arrest; cell cycle G1 phase; concentration response; controlled study; drug cytotoxicity; drug screening; drug structure; enzyme activity; flow cytometry; human; human cell; molecular docking; NTUB1 cell line; prostaglandin release; protein expression; thrombocyte aggregation inhibition; analogs and derivatives; cell proliferation; cell survival; chemical structure; chemistry; dose response; drug effects; G1 phase cell cycle checkpoint; metabolism; molecular dynamics; thrombocyte aggregation; tumor cell line; Antineoplastic Agents; Cell Line, Tumor; Cell Proliferation; Cell Survival; Chalcone; Cyclooxygenase 1; Dose-Response Relationship, Drug; Flow Cytometry; G1 Phase Cell Cycle Checkpoints; Humans; Molecular Docking Simulation; Molecular Dynamics Simulation; Molecular Structure; Platelet Aggregation; Platelet Aggregation Inhibitors; Reactive Oxygen Speciesjournal article10.1248/bpb.b14-00099249890102-s2.0-84904673676