SMALL MOLECULE SCREENING TO IDENTIFY INHIBITORS OF FILOVIRAL

埃博拉病毒（EBOV）是线状病毒群的一员，可引起人类严重的出血热。先天免疫反应是EBOV的重要靶点，Basler博士的研究小组已经确定了抑制干扰素a/p （IFN a/p）产生的丝状病毒蛋白。EBOV VP35 （eVP35）和MARV VP35 （mVP35）通过靶向rig -l样受体信号通路损害IFN a/p的产生。事实上，VP35 ifn拮抗剂受损的EBOV在小鼠和豚鼠中被减弱，这表明VP35抑制剂可用于治疗EBOV感染。Basler博士和Amarasinghe博士的研究小组还表明，EBOV VP24 （eVP24）和MARV VP40 （mVP40）蛋白抑制IFN a/p和IFND触发的Jak-STAT信号，从而阻断IFN的抗病毒作用。我们假设，针对这些丝状病毒蛋白的小分子可以被识别并开发成治疗和预防药物，有助于对抗丝状病毒的生物威胁。它们可以单独使用以增加感染期间ifn的产生，也可以与治疗性给药ifn或可能与其他治疗剂协同使用。这个多项目中心建立在这些令人兴奋的发现和新的检测开发研究（项目1）的基础上，以鉴定eVP35、mVP35、eVP24和mVP40的抑制剂。在项目2中，Microbiofix将使分析适应HTS格式，并进行筛选，确认，验证和选择性/细胞毒性分析，并与德克萨斯大学西南分校的Ready博士团队合作，进行药物化学优化和毒性/PK/PD研究。筛选活动将对Microbiofix（20万种化合物）和ICCB-L（30万种化合物）筛选文库应用4种HTS测定（eVP35、mVP35、eVP24和mVP40）来鉴定作为IFN激动剂的化合物。项目1将描述引线和优化引线抑制的生化、结构和机制基础。感染性核心将在体外和体内对感染丝状病毒的命中和先导化合物进行评价。由此产生的小分子抑制剂可能会产生新的治疗药物，以提高感染患者的预后，以及化学工具来研究这些丝状病毒蛋白的作用。

Ebola virus (EBOV), a member of the filovirus group, causes severe hemorrhagic fever in humans. The Innate immune response is an important target for EBOV, and Dr. Basler's group has identified filovirus proteins that inhibit interferon-a/p (IFN a/p) production. EBOV VP35 (eVP35) and MARV VP35 (mVP35) impair IFN a/p production by targeting RIG-l-like receptor signaling pathways. In fact, EBOVs with impaired VP35 IFN-antagonism are attenuated in mice and guinea pigs indicating that VP35 inhibitors can be used to treat EBOV infection. Dr. Basler's and Dr. Amarasinghe's groups have also shown that EBOV VP24 (eVP24) and MARV VP40 (mVP40) proteins inhibit Jak-STAT signaling triggered by IFN a/p and IFND, thereby blocking the antiviral effects of IFNs. We hypothesize that small molecules targeting these filoviral proteins can be identified and developed into therapeutic and prophylactic agents that are useful for combating filovirus biothreats. They can be used alone to augment the production of IFNs during infection or synergistically in combination with therapeutically-administered IFNs or possibly with other therapeutic agents. This multi-project Center builds on these exciting findings and new assay development studies (Project 1) to identify inhibitors of eVP35, mVP35, eVP24 and mVP40. In Project 2, Microbiofix will adapt the assays to HTS format and conduct screening, confirmation, validation, and selectivity/cytotoxicity assays, and In conjunction with Dr. Ready's group at UT Southwestern, medicinal chemistry optimization, and toxicity/PK/PD studies. The screening campaign will apply 4 HTS assays (eVP35, mVP35, eVP24 and mVP40) to the Microbiofix (200,000 compounds) and ICCB-L (300,000 compounds) screening libraries to identify compounds that act as IFN agonists. Project 1 will characterize the biochemical, structural and mechanistic basis of inhibition by leads and optimized leads. The infectious core will evaluate the hit and lead compounds against infectious filoviruses In vitro and in vivo. Resulting small molecule Inhibitors may be developed to generate new therapeutic agents to enhance patient outcome in Infection as well as chemical tools to study the role of these filoviral proteins.