Cardiovascular and Metabolic Drug Discovery Unit, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan.
Regenerative Medicine Unit, Takeda Pharmaceutical Company Limited, 26-1 Muraoka-higashi 2-Chome, Fujisawa, Kanagawa, 251-8555, Japan.
Sci Rep. 2017 Jul 20;7(1):6050. doi: 10.1038/s41598-017-06481-9.
Purinergic P2Y receptors, G-protein coupled receptors that primarily couple with Gα-proteins, are activated equipotently by adenosine-5'-triphosphate (ATP) and uridine-5'-triphosphate. Evidence suggests that P2Y agonists make potential drug candidates for the treatment of cardiovascular diseases. However, selective non-nucleotide, small-molecule P2Y agonists have yet to be developed. In this report, we discuss Compound 89, a novel non-nucleotide allosteric P2Y agonist that was active in signal transduction and gene induction, and in our in vitro cardiac hypertrophy model. Compound 89 exhibited selective P2Y agonistic activity and potentiated responses to the endogenous agonist ATP, while exhibiting no agonistic activities for four other Gα-coupled human P2Y (hP2Y) receptors and one representative Gα-coupled hP2Y receptor. Its P2Y agonistic effect on mouse P2Y receptors suggested non-species-specific activity. Compound 89 acted as a pure positive allosteric modulator in a Ca mobilization assay of neonatal rat cardiomyocytes; it potentiated ATP-induced expression of genes in the nuclear receptor 4A family (negative regulators of hypertrophic stimuli in cardiomyocytes). Additionally, Compound 89 attenuated isoproterenol-induced cardiac hypertrophy, presumably through dose-dependent interaction with pericellular ATP. These results indicate that Compound 89 is potentially efficacious against cardiomyocytes and therefore a good proof-of-concept tool for elucidating the therapeutic potential of P2Y activation in various cardiovascular diseases.
嘌呤能 P2Y 受体是 G 蛋白偶联受体，主要与 Gα 蛋白偶联，被腺苷-5'-三磷酸 (ATP) 和尿苷-5'-三磷酸 (UTP) 等同等激活。有证据表明，P2Y 激动剂可能成为治疗心血管疾病的潜在药物候选物。然而，选择性非核苷酸小分子 P2Y 激动剂尚未开发。在本报告中，我们讨论了一种新型非核苷酸变构 P2Y 激动剂化合物 89，它在信号转导和基因诱导中具有活性，并且在我们的体外心肌肥厚模型中也具有活性。化合物 89 表现出选择性 P2Y 激动活性，增强了对内源性激动剂 ATP 的反应，而对其他四种 Gα 偶联的人 P2Y（hP2Y）受体和一种代表性的 Gα 偶联 hP2Y 受体没有激动活性。其对小鼠 P2Y 受体的 P2Y 激动作用表明具有非种属特异性活性。在新生大鼠心肌细胞的 Ca 动员测定中，化合物 89 作为纯正变构调节剂发挥作用；它增强了核受体 4A 家族（心肌细胞中肥厚刺激的负调节剂）基因对 ATP 的诱导表达。此外，化合物 89 减弱了异丙肾上腺素诱导的心肌肥厚，可能是通过与细胞外 ATP 的剂量依赖性相互作用。这些结果表明，化合物 89 可能对心肌细胞有效，因此是阐明各种心血管疾病中 P2Y 激活治疗潜力的良好概念验证工具。
