Repurposing the KCa3.1 Blocker Senicapoc for Ischemic Stroke.
Repurposing the KCa3.1 Blocker Senicapoc for Ischemic Stroke.
复制标题
重新利用 KCa3.1 阻滞剂 Senicapoc 治疗缺血性中风。
DOI：
10.1007/s12975-023-01152-6
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发表时间：
2023
影响因子：
6.9
通讯作者：
Wulff,Heike
中科院分区：
文献类型：
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作者：
Lee,RuthD;Chen,Yi-Je;Nguyen,HaiM;Singh,Latika;Dietrich,ConnorJ;Pyles,BenjaminR;Cui,Yanjun;Weinstein,JonathanR;Wulff,Heike
Senicapoc是一种钙激活钾通道KCa3.1的小分子抑制剂，在镰状细胞贫血的临床试验中安全且耐受性良好。我们之前报道的概念验证数据表明，药理学抑制和KCa3.1的基因缺失通过减轻神经炎症减少啮齿动物的梗死并改善神经恢复。在这里，我们评估了重新利用senicapoc治疗缺血性卒中的潜力。在培养的小胶质细胞中，senicapoc抑制KCa 3.1电流，IC 50为7 nM，减少嘌呤激动剂ATP诱导的Ca 2+信号传导，抑制促炎细胞因子和酶的表达，（iNOS和考克斯-2），并阻止炎症体成分NLRP 3的诱导。在雄性C57 BL/6 J小鼠中诱导（tMCAO，60 min），在再灌注后12小时开始每天两次给予10和40 mg/kg的senicapoc，通过T2加权磁共振成像（MRI）测定，剂量依赖性地减少梗死面积并在第8天改善神经功能缺损。超高效液相色谱/质谱分析的总和游离脑浓度证明了足够的KCa3.1目标接合。Senicapoc治疗显著减少小胶质细胞/巨噬细胞和T细胞浸润和活化，并减弱神经元死亡。在3小时开始使用senicapoc的不同治疗模式，在第3天和第8天进行MRI显示senicapoc减少继发性梗死生长并抑制炎症标志物的表达，包括脑中的T细胞细胞因子。最后，我们证明了senicapoc在体外不损害组织纤溶酶原激活剂（tPA）的蛋白水解活性。我们认为，senicapoc可以作为一种持续的免疫细胞保护剂，与缺血性中风的再灌注治疗相结合。
Senicapoc, a small molecule inhibitor of the calcium-activated potassium channel KCa3.1, was safe and well-tolerated in clinical trials for sickle cell anemia. We previously reported proof-of-concept data suggesting that both pharmacological inhibition and genetic deletion of KCa3.1 reduces infarction and improves neurologic recovery in rodents by attenuating neuroinflammation. Here we evaluated the potential of repurposing senicapoc for ischemic stroke. In cultured microglia, senicapoc inhibited KCa3.1 currents with an IC50of 7 nM, reduced Ca2+signaling induced by the purinergic agonist ATP, suppressed expression of pro-inflammatory cytokines and enzymes (iNOS and COX-2), and prevented induction of the inflammasome component NLRP3.When transient middle cerebral artery occlusion (tMCAO, 60 min) was induced in male C57BL/6 J mice, twice daily administration of senicapoc at 10 and 40 mg/kg starting 12 h after reperfusion dose-dependently reduced infarct area determined by T2-weighted magnetic resonance imaging (MRI) and improved neurological deficit on day 8. Ultra-high-performance liquid chromatography/mass spectrometry analysis of total and free brain concentrations demonstrated sufficient KCa3.1 target engagement. Senicapoc treatment significantly reduced microglia/macrophage and T cell infiltration and activation and attenuated neuronal death. A different treatment paradigm with senicapoc started at 3 h and MRI on day 3 and day 8 revealed that senicapoc reduces secondary infarct growth and suppresses expression of inflammation markers, including T cell cytokines in the brain. Lastly, we demonstrated that senicapoc does not impair the proteolytic activity of tissue plasminogen activator (tPA) in vitro. We suggest that senicapoc could be repurposed as an adjunctive immunocytoprotective agent for combination with reperfusion therapy for ischemic stroke.