p38 MAP kinase inhibitors as therapeutics targeting ebolavirus entry

描述（由申请人提供）：线状病毒，埃博拉病毒和马尔堡病毒（EBOV和MARV），是新兴的，包膜的，负义RNA病毒和NIAID A类优先病原体。目前，还没有批准的抗丝状病毒治疗药物。巨噬细胞和树突状细胞（dc）是体内EBOV感染特别重要的细胞类型。这些是感染的早期目标，支持病毒的生产性复制。有证据表明，这些细胞类型的感染会导致导致EBOV疾病的异常反应。例如，感染的巨噬细胞产生促炎细胞因子，可能促进血管渗漏，感染的dc可能失调，损害对感染的免疫反应。因此，预防这些细胞类型感染或改变它们对感染的反应的治疗方法可能是有益的。初步研究表明，先前表征为p38 MAP激酶（p38 MAPK）抑制剂的小分子化合物在原代人巨噬细胞和dc中损害扎伊尔埃博拉病毒样颗粒（VLPs）的进入并抑制扎伊尔埃博拉病毒的复制。这些数据表明p38 MAP激酶可能在EBOV进入巨噬细胞和dc中发挥重要作用。他们还提示p38 MAP激酶可能作为抗ebov药物有用。除了它们抑制进入和复制的能力外，p38抑制剂作为抗EBOV药物是有趣的，因为它们已被开发为抗炎药。因此，它们也可能在体内抑制病毒诱导的炎症。此外，一些p38抑制剂已经在人体临床试验中取得进展。因此，有许多现成的p38 MAPK抑制剂；关于这些化合物的毒性，药代动力学和药效学有大量的文献可以促进从体外研究到动物功效研究的过渡。本项目将在体外和小鼠实验中进一步评估p38 MAP激酶抑制剂作为抗丝状病毒药物的作用。该研究将首先确定市售的p38 MAP激酶抑制剂在细胞培养和小鼠中的抗病毒功效，并确定是否通过抑制p38 MAPKs来抑制EBOV的进入和复制。其次，它将确定p38 MAP激酶抑制剂在细胞培养和体内对ebov诱导的细胞因子产生的影响。最后，它将验证p38 MAP激酶是巨噬细胞介导的EBOV进入巨噬细胞和树突状细胞所必需的假设。综上所述，这些药物有可能作为抗

DESCRIPTION (provided by applicant): The filoviruses, Ebola and Marburg viruses (EBOV and MARV), are emerging, enveloped, negative-sense RNA viruses and NIAID category A priority pathogens. Currently, there are no approved anti-filovirus therapeutics. Macrophages and dendritic cells (DCs) are cell types particularly important for EBOV infection in vivo. These are early targets of infection that support productive virus replication. Evidence suggests that infection of these cell types leads to aberrant responses that contribute to EBOV disease. For example, infected macrophages produce proinflammatory cytokines that may promote vascular leakage, and infected DCs may be dysregulated, impairing immune responses to infection. Therefore, therapeutics that prevent infection of these cell types or that modify their responses to infection may prove beneficial. Preliminary studies have demonstrated that small molecule compounds previously characterized as p38 MAP kinase (p38 MAPK) inhibitors impair, in primary human macrophages and DCs, entry of Zaire EBOV virus-like particles (VLPs) and inhibit Zaire EBOV replication. These data suggest that p38 MAP kinases may play an important role in EBOV entry into macrophages and DCs. They also suggest that p38 MAP kinases may be useful as anti-EBOV drugs. In addition to their ability to inhibit entry and replication, p38 inhibitors are intriguing as anti- EBOV agents because they have been developed as anti-inflammatory drugs. Therefore, they may also suppress virus-induced inflammation in vivo. In addition, some p38 inhibitors have progressed as far as human clinical trials. Therefore, there are a number of readily available p38 MAPK inhibitors; and there is a substantial literature on the toxicity, pharmacokinetics and pharmcodynamics of these compounds that could facilitate the transition from in vitro studies to proof of principle animal efficacy studies. This project wil further evaluate p38 MAP kinase inhibitors as anti-filoviral agents in vitro and in mice. It will frst define the antiviral efficacy of commercially-available p38 MAP kinase inhibitors in cell culture and in mice and determine whether the inhibition of EBOV entry and replication occurs through inhibition of p38 MAPKs. Second, it will determine the impact of p38 MAP kinase inhibitors on EBOV-induced cytokine production in cell culture and in vivo. Finally, it will test the hypothesis that p38 MAP kinases are required for macropinocytosis-mediated entry of EBOV into macrophages and dendritic cells. At its conclusion, the potential for these drugs to serve as anti- filovirus entry inhibitors and suppressors of viral inflammation will be defined; and new details o the EBOV entry pathway in macrophages and DCs will be revealed. PUBLIC HEALTH RELEVANCE: Ebolaviruses are a public health concern because they are deadly and could be used by terrorists as bioweapons. Currently there are no approved drugs to treat these infections. This project will test p38 MAP kinase inhibitors as anti-Ebolavirus drugs i cell culture and in vivo.