Developing capsid-importin alpha inhibitors for the treatment of VEEV infection

开发用于治疗 VEEV 感染的衣壳输入蛋白 α 抑制剂

• 批准号: 10458779
• 负责人: DMITRI Konstantinovich KLIMOV
• 金额: $ 68.78万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10458779
• 关键词: Acute; Animals; Antiviral Agents; Antiviral Response; Attenuated Vaccines; Behavioral; Binding; Binding Sites; Biochemical; Biological Assay; Biological Availability; Calorimetry; Capsid; Capsid Proteins; Category B pathogen; Cell Nucleus; Cells; Cessation of life; Chemical Structure; Clinical; Complex; Confocal Microscopy; Data; Decision Making; Disease; Entropy; Free Energy; Frequencies; Generations; Genetic Transcription; Half-Life; Herpesvirus 1; Human; Hybrids; In Vitro; Inflammation; Knowledge; Laboratory Personnel; Lead; Libraries; Liver Microsomes; Maps; Measures; Memory Loss; Military Personnel; Modification; Molecular; Mutation; National Institute of Allergy and Infectious Disease; Neurons; Nuclear; Pathology; Patients; Permeability; Pharmaceutical Preparations; Proteins; Resistance development; Risk; Seizures; Series; Structure-Activity Relationship; Symptoms; Testing; Therapeutic; Titrations; Toxic effect; Venezuelan Equine Encephalitis Virus; Viral; Viral Load result; Viral Pathogenesis; Virulence; Virus Diseases; Virus Replication; Western Equine Encephalitis Virus; alpha Karyopherins; antiviral drug development; base; blood-brain barrier permeabilization; computer studies; cytotoxicity; design; dosage; efficacy study; improved; in silico; in vivo; indexing; inhibitor; microscopic imaging; molecular dynamics; mouse model; next generation; novel; novel therapeutics; pharmacokinetics and pharmacodynamics; prevent; protein aminoacid sequence; protein protein interaction; side effect; simulation; small molecule; small molecule inhibitor; solute; therapeutic target; trafficking; water solubility

ABSTRACT Venezuelan equine encephalitic virus (VEEV) infects humans and is classified as a Category B pathogen by NIAID due to its easy dissemination. In humans, VEEV infection can result in inflammation, acute degenerative neuronal changes, behavioral changes, memory loss and seizures, with severe cases resulting in death. There is currently no antiviral therapeutic treatment for patients infected with VEEV. The live attenuated vaccine TC83 can protect against VEEV infection, but due to a high frequency of adverse side effects, its use is limited to military and at-risk laboratory personnel. Therefore, the discovery of new therapeutics is urgently needed. VEEV is able to suppress host transcription by blocking cellular nuclear trafficking at least partially due to its capsid protein forming a complex with the host proteins importinα/β (Impα/β1) and CRM1. Mutation of the nuclear localization sequence (NLS) of capsid results in loss of viral virulence, indicating that the ability of capsid to enter the nucleus is critical for VEEV pathogenesis and a viable target for antiviral therapeutic development. We hypothesize that small molecule inhibitors that interfere with capsid-Impα protein-protein interaction (PPI) will prevent VEEV induced pathologies. We have identified two novel small molecules, 1111684 and G281-1485, which disrupt the ability of VEEV capsid to interact with Impα, leading to altered capsid localization, decreased viral replication and increased survival of the host cell. Here we propose to design and synthesize second- generation capsid-Impα PPI inhibitors with improved potency and bioavailability. To this end, we proposed 4 interlinked aims: Aim 1: In silico design of second-generation capsid-Impα inhibitors; Aim 2: Synthesis and biochemical characterization of second generation capsid-Impα inhibitors; Aim 3: Determine the in vitro selectivity index and bioavailability of capsid:Impα inhibitors; and Aim 4: Determine the PK/PD/Tox of capsid:Impα inhibitors. The new knowledge gained from our study will be applicable to wide-range applications involving capsid-Impα interactions including HSV-1 or eastern and western equine encephalitic viruses.

抽象的 委内瑞拉马脑炎病毒 (VEEV) 感染人类，被分类为 B 类病原体 NIAID 由于其易于传播。在人类中，VEEV 感染可导致炎症、急性退行性病变 神经元变化、行为变化、记忆丧失和癫痫发作，严重者导致死亡。那里 目前尚无针对 VEEV 感染患者的抗病毒治疗方法。减毒活疫苗TC83 可以预防 VEEV 感染，但由于不良副作用发生频率较高，其使用仅限于 军事和高危实验室人员。因此，迫切需要发现新的治疗方法。维埃夫 能够通过阻断细胞核运输来抑制宿主转录，至少部分归因于其衣壳 与宿主蛋白 importinα/β (Impα/β1) 和 CRM1 形成复合物。核突变 衣壳的定位序列（NLS）导致病毒毒力丧失，表明衣壳进入的能力 细胞核对于 VEEV 发病机制至关重要，也是抗病毒治疗开发的可行靶点。我们 假设干扰衣壳-Impα 蛋白-蛋白相互作用 (PPI) 的小分子抑制剂将 预防 VEEV 诱发的病变。我们已经鉴定出两种新型小分子，1111684 和 G281-1485， 这破坏了 VEEV 衣壳与 Impα 相互作用的能力，导致衣壳定位改变，减少 病毒复制并增加宿主细胞的存活率。在这里，我们建议设计和合成第二个- 一代衣壳-Impα PPI 抑制剂，具有改进的效力和生物利用度。为此，我们提出了4 相互关联的目标： 目标 1：第二代衣壳-Impα 抑制剂的计算机设计；目标 2：合成和 第二代衣壳-Impα抑制剂的生化表征；目标 3：确定体外 衣壳：Impα抑制剂的选择性指数和生物利用度；目标 4：确定 PK/PD/Tox 衣壳：Impα抑制剂。从我们的研究中获得的新知识将适用于广泛的应用 涉及衣壳-Impα 相互作用，包括 HSV-1 或东部和西部马脑炎病毒。