Paper Title
Anti-Platelet Activity of a Newly Synthesized Benzimidazolederivative, M3bim: An in Vitro Investigation

Abstract
Benzimidazoles are the bonded heterocyclic ring systems which form an integral part of vitamin B12 and have been attracting numerous researchers all over the world to assess their potential therapeutic significance. In this study, we investigated the in vitro antiplatelet activity of newly synthesized benzimidazole derivative, M3BIMin collagen, thrombin, ADP and epinephrine-inducedwashed human platelets. M3BIM showed concentration dependent inhibitory action against collagen (1�g/ml) and thrombin (0.01U/ml)-induced washed human platelet aggregation. Besides, at a concentration of 60 μM, M3BIM distinctly abolished collagen-induced ATP release and intracellular Ca2+ mobilization. Additionally, this compound attenuated the collagen-induced phosphorylation of p47, a marker of protein kinase C (PKC), p38MAPK, and pJNK. However, M3BIM pretreatment did not inhibitADP (20 μM) and epinephrine (10 μM) inducedplatelet aggregation even at a maximum concentration of 500 μM. M3BIMshowed no effects on thrombin-induced P-selectin expression and αIIbβ3 activation, evidenced by flow cytometry and clot reaction assays, respectively.Moreover, the toxic effects of this compound was evaluated using zebrafish embryo toxicity (ZET) assay, and results shown that all M3BIM has no comparative cytotoxicity within the range of 25-200 �M.Together, this study affords confirmation for the inhibitory effect of M3BIM on collagen and thrombin induced platelet aggregation in vitro. The probable explanation of the observed effects of M3BIM may be the occurrence of 1-imidazolyl moiety at one end with longer chain length and increased number of free OH groups. These outcomes may recommend that M3BIM can be appraised as a prospective benzeimidazole compound for the treatment of platelet activation-related diseases. Key words- platelets, benzimidazole, collagen, thrombin, ADP, epinephrine, p-selectin, MAPKs