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: 10701677
• 负责人: DMITRI Konstantinovich KLIMOV
• 金额: $ 67.98万
• 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/10701677
• 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; Classification; Clinical; Complex; Confocal Microscopy; Data; Decision Making; Disease; 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; 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 委内瑞拉马脑病病毒（VEEV）感染了人类，并被归类为B类病原体 Niaid由于其易于传播而引起的。在人类中，VEEV感染会导致感染，急性退化 神经元的变化，行为变化，记忆力丧失和癫痫发作，严重病例导致死亡。那里 目前尚未针对感染VEEV的患者进行抗病毒治疗。活疫苗TC83的活疫苗 可以预防VEEV感染，但由于不良副作用的频率很高，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剂。 Veev 能够通过阻止细胞核运输至少部分地抑制宿主转录 蛋白质与宿主蛋白importinα/β（IMPα/β1）和CRM1形成复合物。核突变 衣壳的定位序列（NLS）导致病毒病毒的丧失，表明衣壳进入的能力 该细胞核对于VEEV发病机理至关重要，也是抗病毒疗法发育的可行靶标。我们 假设小分子抑制剂干扰辣椒IMPα蛋白 - 蛋白质相互作用（PPI）将会 预防VEEV诱导的病理。我们已经确定了两个新型的小分子，1111684和G281-1485， 这破坏了Veev Capsid与IMPα相互作用的能力，从而改变了衣壳定位，改善了 病毒复制和增加的宿主细胞存活率。在这里，我们建议设计和合成第二 具有提高效力和生物利用度的capsid-impαPPI抑制剂。为此，我们提出了4 相互关联的目的：目标1：在第二代Capsid-Impα抑制剂的硅设计中；目标2：合成和 第二代CAPSID-IMPα抑制剂的生化表征；目标3：确定体外 CAPSID的选择性指数和生物利用度：IMPα抑制剂；和目标4：确定PK/PD/TOX的 衣壳：IMPα抑制剂。从我们的研究中获得的新知识将适用于大型应用程序 涉及带囊化的IMPα相互作用，包括HSV-1或东马和西马脑脑病毒。

项目成果

Binding of Venezuelan Equine Encephalitis Virus Inhibitors to Importin-α Receptors Explored with All-Atom Replica Exchange Molecular Dynamics.

用全原子复制品交换分子动力学探索委内瑞拉马脑炎病毒抑制剂与输入α受体的结合。

• DOI: 10.1021/acs.jpcb.3c00429
• 发表时间: 2023
• 期刊: The journal of physical chemistry. B
• 作者: Delfing,BryanM;Olson,Audrey;Laracuente,XavierE;Foreman,KennethW;Paige,Mikell;Kehn-Hall,Kylene;Lockhart,Christopher;Klimov,DmitriK
• 通讯作者: Klimov,DmitriK

Can Free Energy Perturbation Simulations Coupled with Replica-Exchange Molecular Dynamics Study Ligands with Distributed Binding Sites?

• DOI: 10.1021/acs.jcim.3c00631
• 发表时间: 2023-08-02
• 期刊: JOURNAL OF CHEMICAL INFORMATION AND MODELING
• 影响因子: 5.6
• 作者: Lockhart，Christopher;Luo，Xingyu;Klimov，Dmitri K.
• 通讯作者: Klimov，Dmitri K.