p38 MAP kinase inhibitors as therapeutics targeting ebolavirus entry

描述（由申请方提供）：丝状病毒、埃博拉病毒和马尔堡病毒（EBOV和MARV）是新出现的、有包膜的、负义RNA病毒和NIAID A类优先病原体。目前，没有批准的抗丝状病毒治疗剂。巨噬细胞和树突状细胞（DC）是对体内EBOV感染特别重要的细胞类型。这些是支持生产性病毒复制的感染早期目标。有证据表明，这些细胞类型的感染导致EBOV疾病的异常反应。例如，受感染的巨噬细胞产生促炎细胞因子，可能促进血管渗漏，受感染的DC可能失调，损害对感染的免疫应答。因此，预防这些细胞类型感染或改变其对感染的反应的治疗剂可能证明是有益的。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EBOV病毒样颗粒（VLP）的进入并抑制扎伊尔EBOV复制。这些数据表明，p38 MAP激酶可能在EBOV进入巨噬细胞和DC中起重要作用。他们还表明，p38 MAP激酶可用作抗EBOV药物。除了它们抑制进入和复制的能力之外，p38抑制剂作为抗EBOV剂是令人感兴趣的，因为它们已经被开发为抗炎药。因此，它们还可以抑制体内病毒诱导的炎症。此外，一些p38抑制剂已经进展到人类临床试验。因此，有许多容易获得的p38 MAPK抑制剂;并且有大量关于这些化合物的毒性、药代动力学和药效学的文献，这些文献可以促进从体外研究过渡到原则性动物疗效研究。本项目将进一步评价p38 MAP激酶抑制剂作为体外和小鼠抗丝状病毒药物的作用。它将首先确定市售p38 MAP激酶抑制剂在细胞培养物和小鼠中的抗病毒功效，并确定是否通过抑制p38 MAPK来抑制EBOV进入和复制。第二，它将确定p38 MAP激酶抑制剂对EBOV诱导的细胞因子在细胞培养和体内产生的影响。最后，它将测试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需的假设。结论是，这些药物作为抗- 将定义丝状病毒进入抑制剂和病毒炎症抑制剂;并将揭示巨噬细胞和DC中EBOV进入途径的新细节。

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.