title: Novel synthetic benzimidazole-derived oligosaccharide, M3BIM, prevents ex vivo platelet aggregation and in vivo thromboembolism.
abstract: BACKGROUND
  Thrombus formation, a phenomenon primarily related to increased platelet activation, plays a key role in cardiovascular and cerebrovascular diseases. Although the established antiplatelet agents, such as aspirin and clopidogrel, have been shown to be beneficial in treating thromboembolic diseases, they have considerable limitations. Hence, the development of more effective and safe antithrombotic agents is necessary to satisfy a substantial unmet clinical need. In recent years, the favorable properties of imidazole-related drugs have prompted medicinal chemists to synthesize numerous novel therapeutic agents. The chemical structure of the benzimidazole backbone has proven antiplatelet properties. Moreover, synthetic oligosaccharides have exhibited antiplatelet properties. Therefore, we developed a new aldo-benzimidazole-derived oligosaccharide compound, M3BIM, for achieving a stronger antiplatelet effect than the drugs which are being used in clinical aspects. We investigated the effects of M3BIM on platelet activation ex vivo and its antithrombotic activity in vivo.
  
  
  RESULTS
  M3BIM (10-50 μM) exhibited a more potent activity in inhibiting platelet aggregation stimulated by collagen than it did in inhibiting that stimulated by thrombin in washed human platelets. The M3BIM treatment revealed no cytotoxicity in zebrafish embryos, even at the highest concentration of 100 μM. In addition, M3BIM inhibited the phosphorylation of phospholipase Cγ2, protein kinase C (PKC), and mitogen-activated protein kinases (MAPKs; extracellular signal-regulated kinase 2 and c-Jun N-terminal kinase 1), and markedly reduced the ATP-release reaction and intracellular calcium mobilization in collagen-activated platelets. By contrast, M3BIM showed no effects on either collagen-induced p38 MAPK and Akt phosphorylation or phorbol 12, 13-dibutyrate-induced PKC activation and platelet aggregation. Moreover, the M3BIM treatment substantially prolonged the closure time in human whole blood, and increased the occlusion time in mesenteric microvessels and attenuated cerebral infarction in mice. For the study of anticoagulant activities, M3BIM showed no significant effects in the prolongation of activated partial thromboplastin time and prothrombin time in mice.
  
  
  CONCLUSIONS
  The findings of our study suggest that M3BIM is a potential therapeutic agent for preventing or treating thromboembolic disorders.
authors.affiliation: Graduate Institute of Medical Sciences and Department of Pharmacology, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 110, Taiwan.
authors.forename: Ting-Lin
authors.initials: TL
authors.lastname: Yen
authors.affiliation: Graduate Institute of Medical Sciences and Department of Pharmacology, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 110, Taiwan.|Department of Obstetrics and Gynecology, Chi-Mei Medical Center, Tainan, Taiwan.
authors.forename: Ming-Ping
authors.initials: MP
authors.lastname: Wu
authors.affiliation: Department of Medical Research and Translational Laboratory, Research Department, Taipei Medical University Hospital, 252 Wu-Hsing St., Taipei, 110, Taiwan.
authors.forename: Chi-Li
authors.initials: CL
authors.lastname: Chung
authors.affiliation: Genomics Research Center, Academia Sinica, Taipei, Taiwan.
authors.forename: Wen-Bin
authors.initials: WB
authors.lastname: Yang
authors.affiliation: Graduate Institute of Medical Sciences and Department of Pharmacology, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 110, Taiwan.
authors.forename: Thanasekaran
authors.initials: T
authors.lastname: Jayakumar
authors.affiliation: Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli, Tamil Nadu, India.
authors.forename: Pitchairaj
authors.initials: P
authors.lastname: Geraldine
authors.affiliation: Department of Biochemistry, College of Medicine, Taipei Medical University, Taipei, Taiwan.
authors.forename: Chih-Ming
authors.initials: CM
authors.lastname: Chou
authors.affiliation: Hemophilia Center, Department of Pediatrics, Taipei Medical University Hospital, Taipei, Taiwan.
authors.forename: Chia-Yau
authors.initials: CY
authors.lastname: Chang
authors.affiliation: Graduate Institute of Medical Sciences and Department of Pharmacology, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 110, Taiwan. 144106@h.tmu.edu.tw.|Department of Medical Research and Translational Laboratory, Research Department, Taipei Medical University Hospital, 252 Wu-Hsing St., Taipei, 110, Taiwan. 144106@h.tmu.edu.tw.
authors.forename: Wan-Jung
authors.initials: WJ
authors.lastname: Lu
authors.affiliation: Graduate Institute of Medical Sciences and Department of Pharmacology, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 110, Taiwan. sheujr@tmu.edu.tw.
authors.forename: Joen-Rong
authors.initials: JR
authors.lastname: Sheu
mesh_terms: D000818:Animals; D001562:Benzimidazoles / Q000737:chemistry / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D051379:Mice; D010974:Platelet Aggregation / Q000187:drug effects*; D013923:Thromboembolism / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D014312:Trisaccharides / Q000737:chemistry / Q000494:pharmacology; D015027:Zebrafish
chemical_list: D001562:Benzimidazoles; D014312:Trisaccharides; C008317:maltotriose; C031000:benzimidazole