基于结构的EV71病毒靶向抑制剂研究

Enterovirus type 71 is one of the pathogens that cause Hand-Foot-Mouse diseases of infants and children. There are no vaccine or antiviral drugs available against EV71. Recently, the 3-dimential crystal structure of EV71 virion was resolved, which paved way for the research of anti-EV71 drugs with high efficiency and low toxicity. The 3D structure revealed the existence of a hydrophobic pocket on VP1, which can affect virus replication, making an ideal target for the design of antivirals. Previously, we predicted the EV71 VP1 3D structure by homology modeling, and designed and synthesized a series of compounds with the core of phenoxyalkyl pyrazine. Using an in vitro screening model, we obtained among these compounds several hits with ideal antiviral efficiency.This research is based on the recently resolved 3D structure of EV71 virion. With the assistance of computer, we aim to design a series of new compounds exactly targeting the pocket on VP1. After successful synthesis of the compounds, a series of in vitro and in vivo evaluation will be carried out to evaluate the antiviral efficiency against EV71. The final goal of this research is to obtain lead compounds with high anti-EV71 efficiency.

肠道病毒71型（EV71）是婴幼儿手足口病的重要病原体之一，其感染可至脑炎等严重神经系统疾病。目前临床上尚无针对 EV71 的有效疫苗和安全高效的抗病毒药物。近期对EV71病毒颗粒三维结构的成功解析为研究具有高度特异性、低毒性的抗EV71药物提供了理论基础。其三维结构显示，在衣壳蛋白VP1上存在一个狭长的疏水口袋，能够影响病毒复制，是进行抗病毒药物设计的理想靶点。在前期工作中，我们基于EV71 VP1蛋白的同源模建数据，设计并合成了以苯氧烷基吡嗪为母核的系列化合物，并利用体外筛选模型筛选出具有良好抗病毒活性的先导化合物。本研究从最新解析的EV71病毒颗粒三维结构入手，通过计算机辅助药物设计的手段，设计针对VP1蛋白疏水口袋的新结构化合物；对上述化合物进行合成后，利用已经建立的体外及体内筛选模型进一步对其抗病毒活性进行评价，最终希望获得具有高效抗病毒活性的先导化合物。

本课题按照研究内容的要求，基于VP1蛋白三维结构完成了新结构化合物的设计、合成以及生物学评价，发现数个化合物具有良好的抗EV71活性，其中一些化合物对CAV16病毒同样具有良好的抗病毒活性。构效分析表明，该系列化合物苯氧基的供电子基团取代对于提高其抗EV71活性有重要作用，而p-单置换苯环的空间位阻与其抗CAV16的活性相关。本研究还对EV71 3C蛋白酶开展了一系列研究，并开展了EV71的抗病毒新机制研究，发现一类新机制化合物对多种病毒具有广谱抗病毒活性，其中对EV71具有高效的体内治疗效果，2mg/kg的给药剂量即可保护小鼠免受致死剂量的病毒攻击。