帕金森病(PD)发病机制复杂，目前尚无有效治疗手段。靶向α-突触核蛋白(α-Syn)和线粒体保护多巴胺能(DA)神经元是抗PD研究热点。前期我们发现创新药物硝酮嗪(TBN)可保护PD动物模型DA神经元，并降低PD转基因小鼠α-Syn水平，延缓运动行为恶化；TBN可激活PGC-1α/Nrf2通路进而改善线粒体功能。研究报道激活Nrf2能促进蛋白降解系统清除α-Syn。TBN如何激活PGC-1α，上调Nrf2从而清除α-Syn，以及TBN有无直接作用靶点蛋白是本项目需解决的关键科学问题。本项目拟采用细胞和转基因小鼠为研究对象，通过分子药理学、染色质免疫共沉淀、病毒介导的基因长效抑制等手段，研究TBN清除α-Syn和改善线粒体功能的机制；通过药物亲和靶点稳定性(DARTS)技术揭示TBN的直接结合蛋白靶点。本项目有望阐明TBN治疗PD的分子机理，为研发具有自主知识产权抗PD新药提供科学依据。

The pathogenesis of Parkinson’s disease (PD) is complicated. Currently there is no effective treatment for PD. Targeting α-synuclein and mitochondrial dysfunction to protect dopaminergic neurons is the hotspot of PD research. Tetramethylpyrazine nitrone (TBN), a novel derivative of tetramethylpyrazine, significantly protects DA neurons in MPTP/6-OHDA induced rodent models of PD; lowers α-synuclein protein level in human A53T α-synuclein transgenic mice and delays the deterioration of motor behavior. Moreover, TBN can activate PGC-1α/Nrf2 signaling pathway and improve mitochondrial dysfunction. Recent studies revealed that activation of Nrf2 promoted the clearance of α-synuclein via protein degradation systems. We hypothesize that TBN protects dopaminergic neuron through down-regulate α-synuclein and improvement of mitochondrial dysfunction via activating PGC-1α/Nrf2 signaling pathways. How does TBN activate PGC-1α and lower α-synuclein? What're the direct binding targets of TBN? In this proposal, we will use the techniques including molecular pharmacology, chromatin immunoprecipitation (ChIP), animal behavioral tests and molecular genetics to elucidate the mechanisms underlying neuroprotection of TBN. We will also use DARTS (drug affinity-responsive target stability) method to identify the direct binding proteins with TBN. The accomplishment of this project would be expected to uncover the novel molecular mechanisms of TBN for treatment of PD, and provide solid support for developing safe and effective anti-PD drug with intellectual property protection.