Department of Molecular Biology , Sejong University , Seoul 05006 , Republic of Korea.
Neural Injury Research Laboratory , University of Ulsan College of Medicine , Seoul 138-736 , Republic of Korea.
ACS Chem Neurosci. 2019 May 15;10(5):2345-2354. doi: 10.1021/acschemneuro.8b00654. Epub 2019 Feb 22.
We recently reported that AMP-activated protein kinase (AMPK) contributes to zinc-induced neuronal death by inducing Bim, a pro-apoptotic Bcl-2 homology domain 3-only protein, in a liver kinase B1 (LKB1)-dependent manner. Current data suggest AMPK plays key roles in excitotoxicity and ischemic brain injury, with zinc neurotoxicity representing at least one mechanism of ischemic neuronal death. Inhibition of AMPK could be a viable therapeutic strategy to prevent ischemic brain injury following stroke. This prompted our search for novel inhibitors of AMPK activity and zinc-induced neuronal death using cultured mouse cortex and a rat model of brain injury after middle cerebral artery occlusion (MCAO). In structure-based virtual screening, 118 compounds were predicted to bind the active site of AMPK α2, and 40 showed in vitro AMPK α2 inhibitory activity comparable to compound C (a well-known, potent AMPK inhibitor). In mouse cortical neuronal cultures, 7 of 40 compound reduced zinc-induced neuronal death at levels comparable to compound C. Ultimately, only agents 2G11 and 1H10 significantly attenuated various types of neuronal death, including oxidative stress, excitotoxicity, and apoptosis. When administered as intracerebroventricular injections prior to permanent MCAO in rats, 2G11 and 1H10 reduced brain infarct volumes, whereas compound C did not. Therefore, these novel AMPK inhibitors could be drug development candidates to treat stroke.
我们最近报道，AMP 激活的蛋白激酶 (AMPK) 通过诱导促凋亡 Bcl-2 同源结构域 3 仅蛋白 Bim，以 LKB1 依赖性方式促进锌诱导的神经元死亡。目前的数据表明，AMPK 在兴奋性毒性和缺血性脑损伤中发挥关键作用，锌神经毒性至少是缺血性神经元死亡的一种机制。抑制 AMPK 可能是预防中风后缺血性脑损伤的可行治疗策略。这促使我们使用培养的小鼠皮层和大脑中动脉闭塞 (MCAO) 后的大鼠模型寻找新型 AMPK 活性和锌诱导的神经元死亡抑制剂。在基于结构的虚拟筛选中，预测有 118 种化合物可与 AMPKα2 的活性部位结合，其中 40 种化合物显示出与化合物 C（一种众所周知的强效 AMPK 抑制剂）相当的体外 AMPKα2 抑制活性。在小鼠皮质神经元培养物中，40 种化合物中有 7 种可降低锌诱导的神经元死亡，其水平与化合物 C 相当。最终，只有试剂 2G11 和 1H10 显著减轻了各种类型的神经元死亡，包括氧化应激、兴奋性毒性和细胞凋亡。在大鼠永久性 MCAO 之前通过脑室内注射给予时，2G11 和 1H10 可减少脑梗死体积，而化合物 C 则不能。因此，这些新型 AMPK 抑制剂可能是治疗中风的药物开发候选物。
