基于桃柘酚分子探针研究桃柘酚对缺血性脑卒中的神经保护作用及作用靶标

缺血性脑卒中是严重危害人类健康的重大疾病之一。缺血性脑卒中的药物治疗已取得了一定的进展，但由于其病理生理机制复杂，所以寻找新的神经保护药物仍是目前脑卒中药物开发研究的热点。通过建立与脑缺血病理过程密切相关的谷氨酸诱导的原代神经细胞损伤的体外模型，随机筛选化合物库，我们得到一个小分子天然产物桃柘酚。桃柘酚具有很好的抗菌活性，但其神经保护活性未见报道。本项目将主要研究桃柘酚在体外神经细胞损伤模型和在体内大鼠局灶性脑缺血模型中的神经保护作用；在初步构效关系研究的基础上，应用基于活性的蛋白质谱学（ABPP）研究方法，结合光亲合标记和环加成反应策略，设计并合成桃柘酚的ABPP探针分子。应用桃柘酚探针分子的标记实验，一方面将发现桃柘酚的作用靶点，为神经保护药物的开发提供新的药物靶标；另一方面阐明桃柘酚神经保护作用的具体分子机制，为桃柘酚类衍生物开发成为新型的神经保护药物提供临床前的实验依据和理论基础。

Ischemic stroke has become one of the serious diseases threatening the health of human being. Medications used for the treatment of ischemic stroke has been made some progress, but due to the complicated pathophysiologic mechanisms of ischemic stroke disease, searching for novel neuroprotective drugs would be a trend for the drug development on ischemic stroke. Using the established in vitro model on glutamate-induced primary neuronal injury, which is closely associated with pathologic process of ischemic stroke, to randomly screen a compound library, we obtained a small-molecule natural product totarol. Totarol has been reported to have anti-microbial activity, but its neuroprotective role has not been reported so far. This project will focus on study of the neuroprotective role of totarol in in vitro neuronal injury models and in in vivo rat ischemic stroke model, and its molecular mechanisms. Based on preliminary structure-activity relationship study, using the activity-based protein profiling (ABPP) method combined with photoaffinity labeling and click chemistry, we will design and synthesize totarol’s ABPP probe, and use it to label target proteins. With this molecular probe of totarol, we will discover the possible protein target directly interacting with totarol, which will provide a new drug target for the neuroprotective drug development, and elucidate a specific molecular mechanism of totarol neuroprotection, which will provide a preclinical experimental basis and theoretical basis for the totarol development into a neuroprotective drug.