Modified diadenosine tetraphosphates with dual specificity for P2Y1 and P2Y12 are potent antagonists of ADP-induced platelet activation
Modified diadenosine tetraphosphates with dual specificity for P2Y1 and P2Y12 are potent antagonists of ADP-induced platelet activation
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DOI：
10.1111/jth.12035
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发表时间：
2012-12-01
影响因子：
10.4
通讯作者：
Frelinger, A. L., III
中科院分区：
文献类型：
--
作者：
Chang, H.;Yanachkov, I. B.;Frelinger, A. L., III
Chang H、Yanachkov IB、Dix EJ、Li YF、Barnard MR、Wright GE、Michelson AD、Frelinger AL 第 3 名。对 P2Y1 和 P2Y12 具有双重特异性的修饰二腺苷四磷酸是 ADP 诱导的血小板活化的有效拮抗剂。血栓栓塞杂志，2012； 10：257380。摘要。背景：二腺苷 5',5'''-P1,P4-四磷酸 (Ap4A) 是一种储存在血小板致密颗粒中的天然化合物，可抑制 ADP 诱导的血小板聚集。 Ap4A 抑制血小板 ADP 受体 P2Y1 和 P2Y12，是 P2Y12 的部分激动剂，并且是血小板 ATP 门控离子通道 P2X1 的完全激动剂。 Ap4A 四磷酸主链的修饰增强了对 ADP 诱导的血小板聚集的抑制。然而，这些Ap4A类似物对人血小板P2Y1、P2Y12和P2X1的影响尚不清楚。目的：测定二腺苷四磷酸类似物对 P2Y1、P2Y12 和 P2X1 的激动剂和拮抗剂活性。方法：我们合成了以下Ap4A类似物：P1,P4-二硫代四磷酸； P2,P3-氯亚甲基四磷酸酯； P1-硫代-P2,P3-氯亚甲基四磷酸酯；和P1，P4-二硫代-P2，P3-氯亚甲基四磷酸盐。然后我们测量了这些类似物对以下方面的影响： (i) ADP 诱导的血小板聚集； (ii) P2Y1 介导的胞质 Ca2+ 变化； (iii) P2Y12 介导的血管舒张剂刺激的磷蛋白磷酸化的变化； (iv) P2X1 介导的细胞外 Ca2+ 进入。结果：磷酸主链经过修饰的 Ap4A 类似物抑制 P2Y1 和 P2Y12，并且对这些受体没有显示出激动剂活性。二硫代修饰增加了对P2Y1、P2Y12和血小板聚集的抑制，而氯亚甲基修饰增加了对P2Y12和血小板聚集的抑制，但降低了P2Y1抑制。结合二硫代和氯亚甲基修饰增强了 P2Y1 和 P2Y12 抑制。与Ap4A相比，每种修饰都降低了对P2X1的激动剂活性，并且双重修饰完全消除了P2X1激动剂活性。结论：与 Ap4A 相比，Ap4A 的四磷酸骨架类似物对 P2X1 的活性降低，但抑制 P2Y1 和 P2Y12，并且更有效地抑制 ADP 诱导的血小板聚集。因此，具有双重受体选择性的二腺苷四磷酸类似物可能具有作为抗血小板药物的潜力。
Chang H, Yanachkov IB, Dix EJ, Li YF, Barnard MR, Wright GE, Michelson AD, Frelinger AL 3rd. Modified diadenosine tetraphosphates with dual specificity for P2Y1 and P2Y12 are potent antagonists of ADP-induced platelet activation. J Thromb Haemost 2012; 10: 257380. Summary. Background: Diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A), a natural compound stored in platelet dense granules, inhibits ADP-induced platelet aggregation. Ap4A inhibits the platelet ADP receptors P2Y1 and P2Y12, is a partial agonist of P2Y12, and is a full agonist of the platelet ATP-gated ion channel P2X1. Modification of the Ap4A tetraphosphate backbone enhances inhibition of ADP-induced platelet aggregation. However, the effects of these Ap4A analogs on human platelet P2Y1, P2Y12 and P2X1 are unclear. Objective: To determine the agonist and antagonist activities of diadenosine tetraphosphate analogs towards P2Y1, P2Y12, and P2X1. Methods: We synthesized the following Ap4A analogs: P1,P4-dithiotetraphosphate; P2,P3-chloromethylenetetraphosphate; P1-thio-P2,P3-chloromethylenetetraphosphate; and P1,P4-dithio-P2,P3-chloromethylenetetraphosphate. We then measured the effects of these analogs on: (i) ADP-induced platelet aggregation; (ii) P2Y1-mediated changes in cytosolic Ca2+; (iii) P2Y12-mediated changes in vasodilator-stimulated phosphoprotein phosphorylation; and (iv) P2X1-mediated entry of extracellular Ca2+.Results: Ap4A analogs with modifications in the phosphate backbone inhibited both P2Y1 and P2Y12, and showed no agonist activity towards these receptors. The dithio modification increased inhibition of P2Y1, P2Y12, and platelet aggregation, whereas the chloromethylene modification increased inhibition of P2Y12 and platelet aggregation, but decreased P2Y1 inhibition. Combining the dithio and chloromethylene modifications increased P2Y1 and P2Y12 inhibition. As compared with Ap4A, each modification decreased agonist activity towards P2X1, and the dual modification completely eliminated P2X1 agonist activity. Conclusions: As compared with Ap4A, tetraphosphate backbone analogs of Ap4A have diminished activity towards P2X1 but inhibit both P2Y1 and P2Y12 and, with greater potency, inhibit ADP-induced platelet aggregation. Thus, diadenosine tetraphosphate analogs with dual receptor selectivity may have potential as antiplatelet drugs.