Comparative Studies on the Antiplatelet Effects of a Humanized Anti-Platelet Glycoprotein IIb/IIIa Antibody (YM337) and ReoPro under Flow Conditions

Summary

Recent clinical studies have shown that the interruption of platelet function appears to be effective for treatment of coronary occlusive diseases. For this purpose, a Fab fragment of humanized anti-platelet glycoprotein (GP) IIb/IIIa monoclonal antibody (YM 337) that exhibited an in vivo antithrombotic effect without prolongation of bleeding time in monkeys was previously characterized. In this study, the effect of YM 337 under physiological flow conditions with high or low shear rate was evaluated. The antiplatelet effects of YM 337 under varying wall shear rates were examined with the whole blood flow system. This technique allows real time visualization of the formation of fluorescence-labeled platelet thrombi on a collagen surface in a parallel plate flow chamber mounted on an epifluorescence microscope. The process of thrombus growth was also recorded in video tape and subjected to computer-assisted image analysis. We found that YM 337 displayed a high shear-preferential antiplatelet effect, while ReoPro, a control anti-GP IIb/IIIa antibody, did not show such shear dependency. The present findings therefore suggest that YM 337 might be a useful antiplatelet agent which can block pathological thrombotic events occurring under high shear, such as in coronary occlusive diseases.

• References

• 1 Back CH, Radbil JR, Cawford DW. Analysis of pulsatile viscous blood flow through diseased coronary arteries of man.. J Biochem 1977; 10: 339-53.
• 2 Kroll MH, Hellums JD, McIntire LV, Schafer AI, Moake JL. Platelets and shear stress.. Blood 1996; 88: 1525-41.
• 3 Shafer AI. Antiplatelet therapy.. Am J Med 1996; 101: 199-209.
• 4 Topol EJ, Plow EF. Clinical trials of platelet receptor inhibitors.. Thromb Haemost 1993; 70: 94-8.
• 5 EPIC Investigators.. Use of a monoclonal antibody directed against the platelet glycoprotein IIb/IIIa receptor in high risk coronary angioplasty.. N Engl J Med 1994; 330: 956-61.
• 6 Konstantopoulos K, Kamat SK, Shafer AI, Banez EI, Jordan R, Kleiman NS, Hellums ID. Shear-induced platelet aggregation is inhibited by in vivo infusion of an anti-glycoprotein IIb/IIIa antibody fragment, c7E3 Fab, in patients and undergoing coronary angioplasty.. Circulation. 1995; 91: 1427-31.
• 7 Ikeda Y, Handa M, Kawano K, Kamata T, Murata M, Arai Y, Anbo H, Kawai Y, Watanabe K, Iragaki I, Sakai K, Ruggeri ZM. The role of von Willebrand factor and fibrinogen in platelet aggregation under varying shear stress.. J Clin Invest 1991; 87: 1234-40.
• 8 Savage B, Saldivar E, Ruggeri ZM. Initiation of platelet adhesion by arrest onto fibrinogen or translocation on von Willebrand factor.. Cell 1996; 84: 289-97.
• 9 Yano S, Suzuki K, Katoh M, Sugita Y, Kaku S, Kawamura K, Masuho Y. Epitopes and biological activities of two monoclonal antibodies to platelet integrin α IIb β 3.. J Biochem 1994; 116: 778-86.
• 10 Co MS, Yano S, Hsu RK, Landolfi NF, Vasquez M, Cole M, Tso JT, Bringman T, Laird W, Hudson D, Kawamura K, Suzuki K, Furuichi K, Queen C, Masuho Y. A humanized antibody specific for the platelet integrin GP IIb/IIIa.. J Immunol 1994; 152: 2968-76.
• 11 Kaku S, Kawasaki T, Hisamichi N, Sakai Y, Taniuchi Y, Inagaki O, Yano S, Suzuki K, Terazaki C, Masuho Y, Satoh N, Takenaka T, Yanagi K, Oshima N. Antiplatelet and antithrombotic effects of YM 337, the Fab fragment of a humanized anti-GP IIb/IIIa monoclonal antibody in monkeys.. Thromb Haemost 1996; 75: 679-84.
• 12 Aleviadou BR, Moake JL, Turner NA, Ruggeri ZM, Folie BJ, Phillips MD, Schreiber AB, Hrinda ME, McIntire LV. Real-time analysis of shear-dependent thrombus formation and its blockage by inhibitors of von Wille-brand factor binding to platelets.. Blood 1993; 81: 1263-76.
• 13 Turner NA, Moake JL, Kamat SG, Schafer AI, Kleiman NS, Jordan R, McIntire LV. Comparative real-time effects on platelet adhesion and aggregation under flowing conditions of in vivo aspirin, heparin, and monoclonal antibody fragment against glycoprotein IIb-IIIa.. Circulation 1995; 91: 1354-62.
• 14 Usami S, Chen HH, Zhao Y, Chien S, Skalak R. Design and construction of a linear shear stress flow chamber.. Ann Biomed Eng 1993; 21: 77-83.
• 15 Kawasaki T, Sakai Y, Taniuchi Y, Sato K, Maruyama K, Shimizu M, Kaku S, Yano S, Inagaki O, Tomioka K, Yanagisawa I, Takenaka T. Biochemical characterization of a new disintegrin, flavostatin, isolated from Trimeresurus flavoviridis venom.. Biochimie 1996; 78: 245-52.
• 16 Moroi M, Jung SM, Shinmyozu K, Tomiyama Y, Ordinas A, Diaz-Ricart M. Analysis of platelet adhesion to a collagen-coated surface under flow conditions: The involvement of glycoprotein VI in the platelet adhesion.. Blood 1996; 88: 2081-92.
• 17 Kehrel B, Konenberg A, Rauterberg J, Niesing-Bresch D, Nieheus U, Kardoeus J, Schwippert B, Tshope D, van de Loo J, Clemetson KJ. Platelet deficient in glycoprotein IIIb aggregate normally to collagens type I and III but not type V.. Blood 1993; 82: 3364-70.
• 18 Saelman EUM, Nieuwenhuis HK, Hase KM, de Groot PG, Heijnen HFG, Saga EH, Williams S, McKeown L, Gralnick HR, Sixma JJ. Platelet adhesion to collagen type I through VIII under conditions of stasis and flow is mediated by GP Ia/IIa (α 2 β 1-integrin).. Blood 1994; 83: 1244-50.