Receptor-directed small-molecule inhibitors of New World hemorrhagic fever mammarenavirus entry

新世界出血热乳腺病毒入侵的受体定向小分子抑制剂

• 批准号: 10193781
• 负责人: Brian B. Gowen
• 金额: $ 16.02万
• 依托单位: UTAH STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-24 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10193781
• 关键词: Advanced Development; Americas; Animals; Antiviral Agents; Antiviral Therapy; Apical; Binding; Binding Proteins; Biological Availability; Bolivian Hemorrhagic Fever Virus; Cell Culture Techniques; Cells; Chemicals; Complex; Disease; Docking; Dose; Drug Kinetics; Drug resistance; Evaluation; Foundations; Frequencies; GTPBP1 gene; Glycoproteins; Goals; Hemochromatosis; Histopathology; Human; In Vitro; Infection; Investigational Drugs; Junin virus; Knock-in Mouse; Lead; Life; Ligands; Maximum Tolerated Dose; Measures; Metabolic; Metabolism; Modeling; Modification; Mus; National Security; Oral; Pathogenicity; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Plasma Proteins; Population; Property; Proteins; Public Health; Records; Research; Research Project Grants; Risk; Rodent; Science; Severity of illness; Site; Solubility; Structure; Structure-Activity Relationship; Syndrome; TFRC gene; Testing; Therapeutic; Therapeutic Agents; Transferrin; Vaccines; Viral; Viral Hemorrhagic Fevers; Viral Load result; Virus; Zoonoses; absorption; analog; aptamer; base; cytotoxicity; design; drug development; druggable target; efficacy study; experimental study; illness length; improved; indexing; innovation; lead candidate; lead optimization; molecular modeling; mouse model; nanomolar; nonhuman primate; novel; pharmacophore; preclinical development; receptor; scaffold; small molecule inhibitor; small molecule therapeutics; success; survival outcome; therapeutic candidate; therapeutic target; transmission process; treatment duration; virology; virtual screening; virus envelope

PROJECT SUMMARY Mammarenaviruses are endemic in rodent populations worldwide and zoonotic transmission can lead to severe life-threatening hemorrhagic fever. In the Americas, five mammarenavirus species, including Junín and Machupo viruses (JUNV and MACV, respectively), cause viral hemorrhagic fever. In the absence of FDA-licensed antiviral therapies or vaccines, these viruses pose a significant public health concern and threaten national security. The pathogenic New World mammarenaviruses (NWMs) utilize the human transferrin receptor 1 (hTfR1) for entry into human cells. The GP1 subunit of the virus envelope glycoprotein binds to the apical domain of hTfR1, a region that is not involved in the binding to the major hTfR1 ligands. This interaction between GP1 and hTfR1 therefore represents a potential therapeutic target for broad inhibition of NWM infection. Based on the structure of the MACV envelope glycoprotein GP1 subunit in complex with hTfR1, we identified a novel druggable site in the apical domain of hTfR1. A docking-based virtual screening campaign identified 28 hits that were subsequently characterized to determine antiviral activity against JUNV infection in cell culture. Based on this assessment, we selected two chemically distinct molecules displaying strong activity against JUNV for further consideration. We hypothesize that targeting the apical domain pocket of hTfR1 that interacts with viral GP1 will broadly inhibit infection by all known pathogenic NWMs and thereby protect mice expressing hTfR1 from lethal disease associated with JUNV infection. To explore this hypothesis, we will pursue the following specific aims. Aim 1. Identify potent and broadly active compounds targeting the druggable hTfR1 apical domain site that interacts with NWM GP1. The two distinct chemical scaffolds identified in our preliminary studies will serve as the foundation for the design of analogs to define structure-activity relationships (SARs) important for broad antiviral activity. We will implement an iterative campaign involving molecular modeling, medicinal chemistry and in vitro antiviral testing against native JUNV and pseudotyped viruses displaying the envelope glycoprotein of NWMs. Our strategy will include optimization for drug-like properties. Aim 2. Evaluate pharmacokinetics (PK) and efficacy of optimized lead candidates in hTfR1 mice. Two optimized compounds from each chemical scaffold will be chosen for evaluation in animals based on their in vitro potency, selectivity, breadth of inhibition and drug-like properties. We will assess oral bioavailability, determine the maximum tolerated dose and establish PK profiles for the lead candidates in mice. Results from these studies will guide decisions regarding dosing levels and the frequency and duration of treatment in subsequent efficacy studies in a validated hTfR1 mouse model of lethal JUNV infection. In addition to improved survival outcome in JUNV-challenged hTfR1 mice, we will measure the impact of treatments on viral loads, histopathology, and disease severity and duration. Our long-term goal is to develop a novel host-directed small-molecule therapeutic agent that could be deployed to treat NWM hemorrhagic fevers.

项目摘要 哺乳动物肾病毒是世界范围内啮齿动物种群的地方病，人畜共患传播可导致严重的 危及生命的出血热在美洲，包括胡宁和马丘波在内的五种哺乳动物病毒 病毒（分别为JUNV和MACV）引起病毒性出血热。在缺乏FDA许可的抗病毒药物的情况下 这些病毒对公共卫生构成重大关切，并威胁到国家安全。的 致病性新世界哺乳动物肾病毒（NWM）利用人转铁蛋白受体1（hTfR 1）进入 植入人体细胞病毒包膜糖蛋白的GP 1亚基与hTfR 1的顶端结构域结合， 不参与与主要hTfR 1配体结合的区域。GP 1和hTfR 1之间的相互作用 因此代表了广泛抑制NWM感染的潜在治疗靶点。基于结构 MACV包膜糖蛋白GP 1亚基与hTfR 1复合后，我们发现了一个新的药物位点， hTfR 1的顶端结构域。一项基于对接的虚拟筛选活动确定了28个热门歌曲 随后在细胞培养物中表征以确定针对JUNV感染的抗病毒活性。基于此 评估，我们选择了两种化学上不同的分子，对JUNV显示出强活性，用于进一步研究。 考虑.我们假设靶向与病毒GP 1相互作用的hTfR 1顶端结构域口袋， 将广泛抑制所有已知致病性NWM的感染，从而保护表达hTfR 1的小鼠免受 与JUNV感染相关的致命疾病。为了探索这一假设，我们将探讨以下具体问题： 目标。目标1.鉴定靶向可药用hTfR 1顶端结构域的有效且广泛活性的化合物 与NWM GP 1相互作用的位点在我们的初步研究中确定的两种不同的化学支架将 作为类似物设计的基础，以定义重要的结构-活性关系（SAR）， 广泛的抗病毒活性。我们将实施一项迭代运动，涉及分子建模，药物 针对天然JUNV和显示包膜的假型病毒的化学和体外抗病毒试验 NWM的糖蛋白。我们的策略将包括优化药物样特性。目标2.评价 在hTfRl小鼠中优化的先导候选物的药代动力学（PK）和功效。两个优化 将基于它们的体外效力选择来自每种化学支架的化合物用于在动物中进行评价， 选择性、抑制宽度和药物样性质。我们将评估口服生物利用度，确定 最大耐受剂量，并建立小鼠中先导候选物的PK曲线。这些研究的结果 将指导关于剂量水平以及后续疗效中治疗频率和持续时间的决定 在经验证的致死性JUNV感染的hTfR 1小鼠模型中的研究。除了改善生存结果外， 在JUNV攻击的hTfR 1小鼠中，我们将测量治疗对病毒载量、组织病理学和免疫学的影响。 疾病的严重程度和持续时间。我们的长期目标是开发一种新的宿主导向的小分子治疗药物， 可用于治疗NWM出血热的药剂。