Evaluation and development of E1-TopoI as a target for anti-HPV therapeutics

描述(申请人提供)：人乳头瘤病毒(HPV)是最常见的性传播感染，几乎所有的宫颈癌和肛门癌，以及超过一半的口腔癌都是由它引起的。目前的HPV治疗方法是切除病变或通过免疫干预(咪喹莫特作为免疫刺激剂，或HPV疫苗以防止感染最常见的HPV)。虽然已经开发出针对许多类型病毒的抗病毒药物，但到目前为止还没有真正的抗病毒药物可用于治疗HPV。由于严重依赖免疫系统进行HPV治疗/预防，HPV感染和癌症仍然是艾滋病毒/艾滋病患者的主要问题，即使在HAART治疗后也是如此。一种直接对抗HPV的真正的HPV抗病毒药物将是对抗HPV感染和癌症的重要武器。最近干扰疱疹病毒启动酶-解旋酶相互作用的小分子抑制剂的成功证明了使用这种方法来治疗HPV是合理的。我们已经确定了HPV DNA复制解旋酶E1和人类拓扑异构酶I之间的相互作用，这对HPV基因组复制至关重要。AIM 1将评估一组预测会破坏与拓扑异构酶相互作用的E1突变，以更全面地定义这种相互作用。在AIM 2中，将使用多肽结合来进一步定义相互作用区域，我们的E1-拓扑异构酶相互作用试验将用于筛选多肽和小分子面板，以确定这种相互作用的第一代抑制剂。在目标3中，将使用预测软件、核磁共振、X射线结晶学和SAXS来评估这种相互作用的结构。每个目标都可以独立实现，但来自每个目标的信息可以协同和加强其他目标。在每个目标中采用多种可实现的方法就可以实现这些目标，而不需要在每个目标中采用所有的方法。结果将提供将用于开发第二代抑制剂的化学和结构信息，这些抑制剂将被作为潜在的抗乳头瘤病毒疗法进行研究。

DESCRIPTION (provided by applicant): Human papillomavirus (HPV) is the most common sexually-transmitted infection, and the cause of nearly all cervical and anogenital, and over half of oral cancers. Current HPV treatment is by lesion removal or through immunological intervention (imiquimod as an immune stimulant, or the HPV vaccines to prevent infection of the most common HPVs). While antiviral agents have been developed against many types of viruses, to date no true antivirals are available against HPV. With the heavy reliance on the immune system for HPV treatment/prevention, HPV infections and cancers remain a major problem for HIV/AIDS patients, even after HAART treatment. A true HPV antiviral that acts directly against HPV would be an important weapon against HPV infections and cancers. Recent successes of small molecule inhibitors that interfere with the herpesvirus primase- helicase interaction justify using such an approach against HPV. We have identified an interaction between the HPV DNA replication helicase, E1, and human Topoisomerase I that is vital for HPV genome duplication. Aim 1 will evaluate a panel of E1 mutations predicted to disrupt the interaction with Topoisomerase to more fully define this interaction. In Aim 2 peptide binding will be used to further define the interaction domain, and our E1-Topoisomerase interaction assays will be used to screen panels of peptides and small molecules to identify first-generation inhibitors of this interaction. In Aim 3 the structure of this interaction will be evaluted using predictive software, NMR, X-ray crystallography, and SAXS. Each Aim can be achieved independently, yet information from each synergizes and enhances the others. Multiple achieveable approaches in each Aim allows for attaining the goals without requiring success of all approaches in each Aim. Results will provide chemical and structural information that will be used in developing second-generation inhibitors that will be investigated as potential antiviral therapeutics against papillomaviruses.