基于小RNA病毒2C解旋酶核心结构特征的抗病毒策略

小RNA病毒包括许多人类病毒，曾引发多次疫情爆发，是公共卫生安全的威胁。然而，小RNA病毒药物完全缺失。尽管PV和EV71疫苗上市，仍有许多病毒缺乏疫苗。COVID-19疫情警示：提前进行药物、疫苗的技术储备是应对突发传染病疫情的关键。本项目选择小RNA病毒中进化差距较大的HAV 2C与EV71 2C,一方面解析HAV 2C结构，揭示其活性及在复制中的作用；另一方面与EV71 2C比对，验证C端介导的2C-2C互作是最保守的核心特征。基于该特征设计包含C端Pocket binding Domain的多肽；利用体外体系（SPR, ITC, ATPase assay等）和细胞模型评价多肽对病毒复制的抑制能力；确定1-2种活性较高的候选多肽；解析多肽-2C复合体的结构，揭示作用机理；最终，概念验证干扰2C-2C互作是通用抗病毒策略。申请人具有在2C蛋白研究领域具有长期积累和较强创新能力。

Picornaviridae includes many human viruses, have caused multiple outbreaks and are a threat to public health. However, drugs against Picornavirus are unavailable. Although PV and EV71 vaccines are on the market, there are still many viruses that lack vaccines. COVID-19 Pandemic warns: the technical reserve of drugs and vaccines in advance is the key to rapid response to outbreaks of infectious diseases in future. In this project, we selected HAV 2C and EV71 2C, which evolutionary distant in Picornaviridae. On one hand, we will determine the structure of HAV 2C, and reveal its activity and role in replication; on the other hand, comparing with EV71 2C, we prove that 2C-2C interaction is the most conservative core feature. Based on this feature, we will design peptides containing the C-terminal Pocket binding Domain; using in vitro assays (SPR, ITC, ATPase assay, etc.) and cell-base models, we will evaluate the polypeptides’ ability to inhibit viral replication; 1-2 candidate peptides with high activity will be determined; we will then determine the structure of the polypeptide-2C complex and reveal the mechanism of peptide’s action; finally, we will provide a proof of concept, that interfering 2C-2C interaction is a general anti-virus strategy. The applicant has long-term focus, abundant related research achievements and strong innovation ability in the field of 2C protein research.