Selectivity, efficacy and toxicity studies of UCCB01-144, a dimeric neuroprotective PSD-95 inhibitor
Selectivity, efficacy and toxicity studies of UCCB01-144, a dimeric neuroprotective PSD-95 inhibitor
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DOI：
10.1016/j.neuropharm.2019.02.035
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发表时间：
2019-05-15
影响因子：
4.7
通讯作者：
Stromgaard, Kristian
中科院分区：
文献类型：
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作者：
Bach, Anders;Clausen, Bettina H.;Stromgaard, Kristian
抑制突触后密度蛋白95(PSD-95)可使N-甲基-D-天冬氨酸(NMDA)受体下游信号去偶联，从而发挥脑缺血后的神经保护作用。我们之前已经开发了一种二聚体PSD-95抑制剂UCCB01-144，它与PSD-95高亲和力结合，对实验性卒中具有神经保护作用。在这里，我们考察了UCCB01-144的选择性、有效性和毒性，并与单体候选药物TAT-NR2B9c进行了比较。纯化蛋白的荧光偏振和小鼠脑裂解产物的下拉实验表明，UCCB01-144有效地结合了所有四种PSD-95样膜相关鸟苷酸晚期蛋白激酶(MAGUK)。此外，UCCB01-144还影响缺血脑组织中的NMDA受体信号通路。UCCB01-144在小鼠大脑中动脉永久性闭塞30min后给药，不同剂量的UCCB01-144均能缩小青年和老年雄性小鼠的脑梗塞面积，但对青年雄性小鼠和雌性小鼠无明显作用。此外，UCCB01-144在大鼠短暂性卒中模型中具有神经保护作用，与TAT-NR2B9c相比，高剂量UCCB01-144不会导致平均动脉压或心率发生显著变化。总体而言，UCCB01-144是一种有效的MAGUK抑制剂，可减少神经毒性PSD-95介导的信号转导，并在不同条件下改善啮齿类动物的局灶性脑缺血后神经元存活，而不会引起心血管副作用，这鼓励了对中风临床试验的进一步研究。
Inhibition of postsynaptic density protein-95 (PSD-95) decouples N-methyl-D-aspartate (NMDA) receptor downstream signaling and results in neuroprotection after focal cerebral ischemia. We have previously developed UCCB01-144, a dimeric PSD-95 inhibitor, which binds PSD-95 with high affinity and is neuroprotective in experimental stroke. Here, we investigate the selectivity, efficacy and toxicity of UCCB01-144 and compare with the monomeric drug candidate Tat-NR2B9c. Fluorescence polarization using purified proteins and pull-downs of mouse brain lysates showed that UCCB01-144 potently binds all four PSD-95-like membrane-associated guanylate kinases (MAGUKs). In addition, UCCB01-144 affected NMDA receptor signaling pathways in ischemic brain tissue. UCCB01-144 reduced infarct size in young and aged male mice at various doses when administered 30 min after permanent middle cerebral artery occlusion, but UCCB01-144 was not effective in young male mice when administered 1 h post-ischemia or in female mice. Furthermore, UCCB01-144 was neuroprotective in a transient stroke model in rats, and in contrast to Tat-NR2B9c, high dose of UCCB01-144 did not lead to significant changes in mean arterial blood pressure or heart rate. Overall, UCCB01-144 is a potent MAGUK inhibitor that reduces neurotoxic PSD-95-mediated signaling and improves neuronal survival following focal brain ischemia in rodents under various conditions and without causing cardiovascular side effects, which encourages further studies towards clinical stroke trials.