A Flexible Approach to Inhibiting HIV NCp7

抑制 HIV NCp7 的灵活方法

• 批准号: 9008023
• 负责人: Katherine L Seley-Radtke
• 金额: $ 19.09万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE COUNTY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-05 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9008023
• 关键词: Adverse effects; Antiviral Agents; Area; Automobile Driving; Binding; Biological; Biological Testing; Complex; Computer Simulation; DNA; Drug Targeting; Enzymes; Exhibits; Future; Generations; Genome; Guanosine; HIV; HIV-1; Hand; Health; Highly Active Antiretroviral Therapy; Laboratories; Left; Literature; Modeling; Modification; Molecular Conformation; Multi-Drug Resistance; Mutation; Nucleocapsid Proteins; Nucleosides; Play; Polyproteins; Procedures; Process; Protein Structure Initiative; Proteins; RNA; RNA Binding; Replication-Associated Process; Reporting; Research; Resistance; Resistance development; Reverse Transcription; Role; Route; Series; Specificity; Staging; Structure; Testing; Therapeutic; Titrations; Toxic effect; Transfer RNA; Viral; Virus Replication; Zinc; Zinc Fingers; analog; base; cytotoxicity; design; drug development; flexibility; global health; inhibitor/antagonist; model design; molecular dynamics; mutant; nucleobase; research study; resistant strain; screening; small molecule inhibitor; student training; virtual; zinc-binding protein

 DESCRIPTION (provided by applicant): The extensive use of antiviral drugs has resulted in the emergence of mutant viral strains that are resistant to the currently available nucleoside therapeutics. Using a combination of drugs that target different viral enzymes (termed highly active anti-retroviral therapy or HAART) has helped, however undesirable side effects and toxicity are often prevalent and reports of multidrug resistance are increasing. As a result, the need for new viral enzymatic targets is critical. In that regard, the nucleocapsid protein of HIV offers forth an attractive target. NCp7 is a small zinc-binding protein that has been shown to be critical in both the early and late stages of the HIV-1 replication process. NCp7 (and the NCp7 domain of the Gag precursor polyprotein) has several key roles in HIV replication, including direct participation in RNA genome recognition and packaging, promoting annealing of the tRNA primer to initiate reverse transcription, and facilitating strand transfer during reverse transcription. NCp7 contains two CCHC-type zinc finger domains that play critical RNA-binding roles during replication, and single-atom Cys-to-Ser mutations that disrupt zinc coordination are sufficient to completely block viral replication. Several approaches to designing inhibitors have been tried, most commonly, use of zinc-ejectors, however to date these have exhibited significant toxicity and poor specificity. A problem that has hindered the identification of viable inhibitors is the inherent flexibility of the protein itself. As such, flexible nucleosides, known s "fleximers" designed and developed in the Seley-Radtke laboratory may provide a solution. In that regard, a series of flexible nucleobase ("flexbase") inhibitors of NCp7 have been identified by means of a virtual screening process. The flexbase analogues are in excellent alignment with the central guanosine residue known to be critical to the biologically relevant conformations of NCp7, thus form the basis for this application. Notably, the flexbases should not result in zinc ejection thereby overcoming the problems associated with other approaches.

 描述(申请人提供)：抗病毒药物的广泛使用导致了对目前可用的核苷疗法产生抗药性的突变病毒株的出现。使用针对不同病毒酶的药物组合(称为高效抗逆转录病毒疗法或HAART)有所帮助，但不良副作用和毒性往往普遍存在，多药耐药的报告也在增加。因此，需要新的病毒酶靶标是至关重要的。在这方面，艾滋病毒的核衣壳蛋白提供了一个有吸引力的目标。NCp7是一种小的锌结合蛋白，已被证明在HIV-1复制过程的早期和晚期都是关键的。NCp7(以及Gag前体多聚蛋白的NCp7结构域)在HIV复制中具有几个关键作用，包括直接参与RNA基因组的识别和包装，促进tRNA引物的退火以启动逆转录，以及促进逆转录过程中的链转移。NCp7含有两个CCHC类型的锌指结构域，它们在复制过程中发挥关键的RNA结合作用，而单原子Cys-to-Ser突变破坏了锌的协调，足以完全阻止病毒的复制。已经尝试了几种设计抑制剂的方法，最常见的是使用锌喷射器，然而到目前为止，这些方法表现出显著的毒性和较差的特异性。这个问题阻碍了确定可行的 抑制剂是蛋白质本身固有的灵活性。因此，在塞利-拉德克实验室设计和开发的被称为S的柔性核苷可能提供一种解决方案。在这方面，已经通过虚拟筛选过程确定了一系列NCp7的柔性碱基(“Flexbase”)抑制剂。柔性碱基类似物与已知对NCp7的生物相关构象至关重要的中心鸟苷残基具有很好的一致性，因此形成了这一应用的基础。值得注意的是，柔性底座不应导致锌喷出，从而克服了与其他方法相关的问题。