Anti-norovirus protease inhibitors for immunocompromised patients

项目总结/摘要 人类诺如病毒属杯状病毒科，是病毒性胃肠炎的主要病原 和食源性疾病。每年仅在美国，hNoV就负责超过 2100万例急性胃肠炎，导致每年约71，000例住院治疗。虽然大多数 病例在一周内消退，免疫功能低下的患者，儿童和老年人的风险增加， 长期甚至致命的感染。目前，还没有抗病毒药物或疫苗被批准用于预防 或治疗慢性hNoV感染。该项目的主要目标是利用医疗福利， 通过抑制病毒酶，包括hNoV蛋白酶来提供。本建议的总体目标是 设计和开发用于治疗hNoV感染的蛋白酶抑制剂。hNoV NS6蛋白酶具有 由于其在病毒复制周期中的重要作用而成为有吸引力的药物靶标。核心假设是 靶向病毒蛋白酶的小分子抑制剂将限制或消除hNoV感染。的理由 所提出的研究是基于我们的初步数据，特别是化合物1及其类似物。这些 化合物是独特的、无毒的、小的肽模拟物分子， 通过与hNoV蛋白酶相互作用以20 nM EC50培养。hNoV蛋白酶的药理学操作 预计将为预防和治疗NoV感染带来新的创新方法。指导 通过强有力的初步数据，这一假设将通过追求三个具体目标来检验：目标1）优化和 通过多参数优化和分子建模发现和开发广谱口服NoV PI 从我们的专有图书馆的铅化合物的方法。目的2）研究其作用机制， 电极导线NoV PI的耐药性研究。目的3）在免疫功能低下的小鼠中评估前导NoV PI 模型在第一个目标下，每年将设计、合成和测试大约25种化合物。下 第二个目的，酶动力学，晶体学，宿主蛋白酶抑制，抑制NoV蛋白酶产生， 并将对先导化合物进行耐药性研究。在第三个目标下，抗NoV功效 将在免疫缺陷小鼠模型中进行研究。这种方法是创新的，因为我们 充分利用五个最新进展制定了研究计划：（1）生产hNoV PR重组体 蛋白质的毫克量;（2）解决的NoV GII.4 PR的晶体结构-主要的药物靶标;（3） 基于细胞的hNoV复制子测定;（4）3D人肠类肠模型，和（5）免疫受损小鼠 疾病模型来研究人类NoV感染。这项研究意义重大，因为深入的临床前研究 这些新的肽模拟物的研究和表征可能导致批准第一个安全且 用于治疗慢性hNoV感染的有效药物， 免疫功能低下的患者。

PROJECT SUMMARY/ABSTRACT Human Noroviruses (hNoV), which belong to the Caliciviridae family, are the leading cause of viral gastroenteritis and food-borne disease worldwide. Each year in the United States alone, hNoV is responsible for greater than 21 million cases of acute gastroenteritis, leading to an estimated 71,000 hospitalizations per year. While most cases resolve within a week, immunocompromised patients, children, and the elderly have an elevated risk of long-term and even fatal infections. Currently, there are no antiviral drugs or vaccines approved for the prevention or treatment of chronic hNoV infections. The main goal of this project is to harness the medical benefits that are offered by the inhibition of viral enzymes, including the hNoV protease. The overall objective of this proposal is to design and develop a protease inhibitor for the treatment of hNoV infections. The hNoV NS6 protease has become an attractive drug target due to its essential role in the viral replication cycle. The central hypothesis is that small-molecule inhibitors targeting the viral protease will limit or eliminate hNoV infections. The rationale for the proposed research is based on our preliminary data, specifically compound 1 and its analogs. These compounds are unique, non-toxic, small peptidomimetic molecules that inhibit NoV replication selectively in culture at 20 nM EC50 by interacting with the hNoV protease. Pharmacological manipulation of the hNoV protease is expected to result in new and innovative approaches to the prevention and treatment of NoV infections. Guided by strong preliminary data, this hypothesis will be tested by pursuing three specific aims: Aim 1) To optimize and discover and develop broad-spectrum oral NoV PIs by multiparameter optimization and molecular modeling approaches of lead compounds from our proprietary library. Aim 2) To perform mechanism of action (MOA) and resistance studies of the lead NoV PIs. Aim 3) To evaluate the lead NoV PIs in an immunocompromised mouse model. Under the first aim, about 25 compounds will be designed, synthesized, and tested per year. Under the second aim, enzyme kinetic, crystallographic, host protease inhibition, inhibition of NoV protease production, and drug resistance studies will be performed with the lead compounds. Under the third aim, anti-NoV efficacy studies will be performed in an immunodeficient mouse model. This approach is innovative because we developed a research plan taking full advantage of five recent advances: (1) producing hNoV PR recombinant proteins in milligram amounts; (2) solving the crystal structure of the NoV GII.4 PR - the main drug target; (3) a cell-based hNoV replicon assay; (4) 3D human intestinal enteroid model, and (5) an immunocompromised mouse disease model to study human NoV infection. The proposed research is significant because in-depth pre-clinical studies and characterization of these new peptidomimetics could lead to the approval of the first safe and effective drugs for the treatment of chronic hNoV infections which cause significant morbidity and mortality in immunocompromised patients.