双靶向PARP1和ULK1小分子设计合成、结构优化和治疗帕金森病作用机制研究

Current clinical drugs against Parkinson’s disease (PD) can improve the symptoms of patients in a short period of time, but the long-term treatment effect is not satisfactory. Recent studies have shown that PARP1 inhibitors block α-synuclein oligomer fibrillization and have great potential for drug development against PD. Considering the cytotoxicity of existing PARP1 inhibitors and the inability to eliminate α-synuclein oligomers completely, the autophagy-initiating key protein ULK1 associated with PARP1 was introduced. The design of small molecule compounds dual-targeting ULK1 and PARP1 could achieve two-pronged treatment of PD by inhibition of α-synuclein oligomerization and promotion of oligomer-clearance. In the previous study, we used computer-aided drug design combing with experiments in vitro and in vivo to discovery the lead compound DT-01, which had biological activities dual-targeting ULK1 and PARP1. The mechanism and efficacy of DT-01 against PD was preliminarily revealed. Based on these results, this project intends to combine the virtual combinatorial library design and free energy evaluation to guide the structural modification and chemical synthesis of DT-01 analogs. Meanwhile, molecular pharmacology and proteomics methods will be applied to study the synergistic mechanism and anti-PD treatment effect of the optimized compounds by regulating signaling pathways dominated by PARP1 and ULK1. All these results would provide a basis for the development of anti-PD innovative drugs from a new strategy.

现有的帕金森病（PD）治疗药物可以在短期内改善患者的症状，但是长期治疗效果并不理想。最新研究表明PARP1抑制剂能阻断α-突触核蛋白寡聚体纤维化，具有抗PD药物开发的巨大潜力。针对已有PARP1抑制剂细胞毒性大、无法清除α-突触核蛋白寡聚体的特点，引入与PARP1相关的自噬启动关键蛋白ULK1，通过双靶向ULK1和PARP1的小分子设计，实现双管齐下治疗PD的目的。我们在前期的研究中，运用计算机辅助药物设计手段，结合体内外实验，发现先导化合物DT-01具有双靶向ULK1和PARP1的活性，初步揭示了DT-01抗PD的机制和药效。本研究拟在此基础上，利用虚拟组合库设计和结合自由能评估来指导DT-01类似物的结构修饰和化学合成。同时，采用分子药理学和蛋白组学的方法深入研究优选化合物对PARP1和ULK1主导信号通路的协同作用机制及抗PD治疗效果，为基于新策略的抗PD创新药物的研发提供基础。

现有的帕金森病（PD）治疗药物缺乏长期有效的治疗效果。PARP1抑制剂具有抗PD药物开发的潜力，但已有PARP1抑制剂细胞毒性较大，通过引入与PARP1相关的自噬启动关键蛋白ULK1，设计双靶向ULK1和PARP1的小分子，有望达到治疗PD的目的。运用计算机辅助药物设计手段和实验评价获得先导化合物DT-01，在此基础上利用组合库设计的思路设计合成了18个小分子化合物。化合物PU5具有较好的PARP1抑制活性，同时在MPP+诱导的SH-SY5Y细胞模型中表现出良好的细胞保护活性。WB实验结果显示PU5对于PARP1、ULK1及相关通路具有调节作用。进一步，在动物体内实验中，各给药组中小鼠的运动障碍均有不同程度的改善，且表现出一定的浓度依赖性；对实验小鼠动物脏器（心、肝、脾、肺、肾）H&E染色结果显示，化合物PU5没有明显的毒性作用，具有一定的安全性。以上结果表明化合物PU5能够靶向调节PARP1和ULK1信号通路，具备较好的抗PD药物开发的潜力。