Discovery and evaluation of anti-filovirus therapeutics targeting NPC1

丝状病毒与偶发性但日益频繁的高度传染性疾病暴发有关， 致命的出血热，是生物恐怖主义的关注因素。没有fda批准 疫苗或药物目前可用于预防或治疗丝状病毒感染。抗病毒药物 靶向病毒感染所必需的宿主成分具有赋予广泛的免疫调节作用的潜力。 对具有类似感染途径的病毒的保护，它们可能不太可能 产生耐药性的药物。不幸的是， 已经鉴定出丝状病毒感染的关键宿主因素。然而，我们最近报道说， 尼曼-匹克C1（NPC 1），一种在所有细胞中表达的溶酶体胆固醇转运蛋白， 丝状病毒广泛需要进入和感染人类细胞，并在 小鼠我们还发现NPC 1中的单个管腔区段（结构域C）介导丝状病毒进入 通过特异性和直接结合丝状病毒GP。因此，GP-NPC 1相互作用 代表了开发广泛活性抗丝状病毒的新的独特分子靶点 治疗学我们的总体目标是确定靶向关键相互作用的小分子 NPC 1与包膜糖蛋白（GP）之间的关系，并将其开发为临床 具有广谱抗丝状病毒活性的候选物。这项提案的主要里程碑是 选择适合抗丝状病毒IND启用的主要和备用临床前候选药物 问题研究为了实现这些目标，PI组建了一个多学科团队，其中包括 公司在抗感染药物的发现和开发方面取得了成功， 病毒学家，在BSL-4下的丝状病毒研究方面具有长期经验 安全壳我们将共同： 1.开发试验以筛选靶向GP-NPC 1相互作用的抑制剂 2.进行高通量化学筛选以鉴定和确认GP-NPC 1相互作用 抑制剂 3.研究了HITS和早期先导化合物的抗病毒活性，并阐明了它们的作用机制 4.针对抗病毒活性和体外药理学对经验证的命中进行化学优化 5.建立铅抑制剂的药理学和毒理学特征，并对其进行评价， 在鼠感染模型中的抗病毒功效

Filoviruses have been associated with episodic, but increasingly frequent outbreaks of a highly lethal hemorrhagic fever, and are agents of concern for bioterrorism. No FDA-approved vaccines or drugs are currently available to prevent or treat filovirus infections. Antiviral drugs targeting host components essential for viral infection have the potential to confer broad protection against viruses with similar infection pathways, and they may be less likely to engender drug resistance than their counterparts targeting viral components. Unfortunately, few host factors critical for filovirus infection have been identified. However, we recently reported that Niemann-Pick C1 (NPC1), a lysosomal cholesterol transporter expressed in all cells, is broadly required by filoviruses for entry and infection of human cells, and for pathogenesis in mice. We also found that a single luminal segment (domain C) in NPC1 mediates filovirus entry by binding specifically and directly to filovirus GP. Therefore, the GP-NPC1 interaction represents a new and unique molecular target for developing broadly active anti-filovirus therapeutics. Our overall goals are to identify small molecules targeting the critical interaction between the envelope glycoprotein (GP) and NPC1, and to develop them into clinical candidates with broad-spectrum activity against filoviruses. The major milestone of this proposal is to select primary and backup pre-clinical candidates suitable for anti-filovirus IND-enabling studies. To fulfill these objectives, the PI has assembled a multidisciplinary team that includes a company with demonstrated success in the discovery and development of anti-infectives and a virologist who brings longstanding experience in the investigation of filoviruses under BSL-4 containment. Together, we will: 1. Develop assays to screen for inhibitors targeting the GP-NPC1 interaction 2. Perform high-throughput chemical screens to identify and confirm GP-NPC1 interaction inhibitors 3. Validate the antiviral activity of hits and early leads and elucidate their mechanisms of action 4. Chemically optimize validated hits for antiviral activity and in vitro pharmacology 5. Establish pharmacologic and toxicologic profiles of lead inhibitors and evaluate them for antiviral efficacy in murine infection models