SMALL MOLECULE SCREENING TO IDENTIFY INHIBITORS OF FILOVIRAL

埃博拉病毒(EBOV)是丝状病毒组的成员，会在人类中引起严重出血热。先天免疫反应是EBOV的一个重要靶点，巴斯勒博士的团队已经确定了抑制干扰素a/p产生的丝状病毒蛋白。EBOV VP35(EVP35)和MARV VP35(MVP35)通过靶向RIG-L样受体信号通路抑制干扰素a/p的产生。事实上，具有受损的VP35干扰素拮抗作用的EBOV在小鼠和豚鼠中得到减弱，这表明VP35抑制剂可以用于治疗EBOV感染。巴斯勒博士和阿马拉辛格博士的团队还表明，EBOV VP24(EVP24)和Marv VP40(MVP40)蛋白抑制由干扰素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，靶向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，对抗击丝状病毒生物治疗有用。它们可以单独用于在感染期间增加干扰素的产生，或与治疗性给药的干扰素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，以确定eVP35、mVP35、eVP24和mVP40的抑制剂。在项目2中，MicroBiofix将使分析方法适应HTS格式，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Dr.Ready团队合作，进行药物化学优化和毒性/PK/PD研究。筛选活动将在MicroBiofix(200,000种化合物)和ICCB-L(300,000种化合物)筛选文库中应用4种HTS分析(eVP35、mVP35、eVP24和mVP40)，以识别作为干扰素激动剂的化合物。项目1将描述铅和优化的铅抑制的生化、结构和机制基础。感染核心将在体外和体内评估抗感染性丝状病毒的HIT和先导化合物。由此产生的小分子抑制剂可能会被开发出来，以产生新的治疗药物，以改善感染患者的预后，以及研究这些丝状病毒蛋白的作用的化学工具。

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.