Identification of a selective small-molecule inhibitor of type 1 adenylyl cyclase activity with analgesic properties
Identification of a selective small-molecule inhibitor of type 1 adenylyl cyclase activity with analgesic properties
复制标题
DOI：
10.1126/scisignal.aah5381
复制
发表时间：
2017-02-21
影响因子：
7.3
通讯作者：
Watts, Val J.
中科院分区：
文献类型：
--
作者：
Brust, Tarsis F.;Alongkronrusmee, Doungkamol;Watts, Val J.
腺酰环化酶1(Ac1)是一组受钙离子刺激的钙调蛋白依赖性腺酰环化酶(Acs)。对AC1基因敲除小鼠的研究表明，AC1的抑制剂可能对治疗疼痛和阿片类药物依赖有用。然而，对AC异构体的非选择性抑制可能会导致严重的不良反应。我们利用化学文库筛选的方法筛选出一种具有色酮核心结构的选择性AC1抑制剂，它可能代表一种新的止痛药。在证明化合物(ST034307)能抑制钙离子刺激的腺苷3‘，5’-单磷酸(CAMP)在稳定转导AC1的人胚胎肾(HEK-AC1)细胞(HEK-AC1细胞)中积聚后，我们通过对膜结合型AC1的所有亚型的检测证实了AC1的选择性。ST034307在HEK-AC1细胞上也能抑制Forskolin和Gas偶联受体刺激的AC1活性，在多种含AC1膜制剂和小鼠海马匀浆中也表现出抑制活性。ST034307增强m-阿片受体(MOR)介导的AC1在稳定表达MOR的HEK-AC1细胞的短程抑制实验中的抑制作用，但该化合物可阻断慢性MOR激活引起的AC1的异源增敏。ST034307可降低炎症性疼痛小鼠模型的疼痛反应。我们的数据表明，ST034307是一种选择性的AC1小分子抑制剂，并提示选择性的AC1抑制剂可能对止痛有用。
Adenylyl cyclase 1 (AC1) belongs to a group of adenylyl cyclases (ACs) that are stimulated by calcium in a calmodulin-dependent manner. Studies with AC1 knockout mice suggest that inhibitors of AC1 may be useful for treating pain and opioid dependence. However, nonselective inhibition of AC isoforms could result in substantial adverse effects. We used chemical library screening to identify a selective AC1 inhibitor with a chromone core structure that may represent a new analgesic agent. After demonstrating that the compound (ST034307) inhibited Ca2+-stimulated adenosine 3',5'monophosphate ( cAMP) accumulation in human embryonic kidney (HEK) cells stably transfected with AC1 (HEK-AC1 cells), we confirmed selectivity for AC1 by testing against all isoforms ofmembrane-bound ACs. ST034307 also inhibited AC1 activity stimulated by forskolin-and Gas-coupled receptors in HEK-AC1 cells and showed inhibitory activity in multiple AC1-containing membrane preparations andmouse hippocampal homogenates. ST034307 enhanced m-opioid receptor (MOR)-mediated inhibition of AC1 in short-terminhibition assays in HEK-AC1 cells stably transfected with MOR; however, the compound blocked heterologous sensitization of AC1 caused by chronic MOR activation in these cells. ST034307 reduced pain responses in a mouse model of inflammatory pain. Our data indicate that ST034307 is a selective small-molecule inhibitor of AC1 and suggest that selective AC1 inhibitors may be useful for managing pain.