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

基本信息

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
项目类别：
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

项目摘要

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 由于其容易在人类中传播，VEEV 感染可导致炎症、急性退行性变。神经变化、行为改变、记忆丧失和癫痫发作，严重者导致死亡。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，其使用仅限于因此，迫切需要发现新的 VEEV 疗法。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。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。接下来，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 引起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，减少病毒复制和增加宿主细胞的存活率在这里我们建议设计和合成第二- 为此，我们提出了 4 种具有改进效力和生物利用度的衣壳-Impα PPI 抑制剂。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；以及目标 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。衣壳-Impα 相互作用，包括 HSV-1 或东部和西部马脑炎病毒。