Hayata Kenji, Nakayama Takayuki, Matsushita Tadashi, Sakano Katsuichi
R&D Division, Exploratory Research Laboratories II, Daiichi-Sankyo Co. Ltd., 16-13 Kita-Kasai 1-Chome, Edogawa-ku, Tokyo, Japan.
J Pharmacol Sci. 2008 Oct;108(2):217-21. doi: 10.1254/jphs.08147sc. Epub 2008 Oct 10.
To obtain compounds that inhibit the interaction of von Willebrand factor (vWF) and glycoprotein (GP) Ib, a novel binding assay was established. The binding of fixed platelets to vWF-R497 mutant was quantified by a solid phase assay. In this assay, fixed platelets bound to the vWF-R497 mutant, carrying the deletion of Glu497-Tyr508 and the missense mutation of Arg545 to Ala, without binding modulators such as ristocetin. The K(d) value of the binding was 2.8 nM, which was consistent with the result from liquid binding assay. The binding was inhibited by aurin tricarboxylic acid (ATA) and an anti GPIb antibody, AK2. Using this binding assay, we screened our library compounds and obtained D74-3736. This compound also inhibited ristocetin-induced platelet aggregation in the human platelet-rich plasma.
为了获得抑制血管性血友病因子（vWF）与糖蛋白（GP）Ib相互作用的化合物，建立了一种新型结合测定法。通过固相测定法定量固定化血小板与vWF-R497突变体的结合。在该测定法中，固定化血小板与携带Glu497-Tyr508缺失以及Arg545突变为Ala的错义突变的vWF-R497突变体结合，无需诸如瑞斯托霉素等结合调节剂。结合的K(d)值为2.8 nM，这与液体结合测定法的结果一致。该结合被金精三羧酸（ATA）和抗GPIb抗体AK2抑制。使用这种结合测定法，我们筛选了我们的文库化合物并获得了D74-3736。该化合物也抑制人富含血小板血浆中瑞斯托霉素诱导的血小板聚集。
