Antiviral Compounds for Human Papillomavirus 16

该项目寻求将咪唑-吡咯聚酰胺用作治疗人乳头瘤病毒(HPV)的局部抗病毒药物。人乳头瘤病毒，特别是以性传播形式传播的人乳头瘤病毒，在世界各地构成了一个重大的公共卫生问题。目前的治疗方法，包括手术、冷冻、有毒物质和生物反应调节剂，都是原始的或非特异性的。缺乏治疗替代方案的一个原因是病毒基因组较小，以及相应地缺乏抗病毒靶点。出于这个原因，人们正在采取使用序列特异性DNA结合剂靶向HPV DNA的替代方法。 HPV复制起点(ON)包含病毒编码的复制蛋白E1和E2的结合位点，这是病毒复制和维护所必需的。靶向或/与序列特异性DNA结合剂有可能阻止E1和E2的作用，从而扰乱细胞维持所需的过程。HPV31的初步研究显示了重大的前景，咪唑-吡咯聚酰胺，旨在靶向HPV31E1结合位点， 被发现能有效地降低细胞中HPV31的外周DNA水平。我们现在请求SUPPED用于针对具有商业重要性的HPV16基因的化合物的设计、合成和测试。 与我们对HPV31的初步研究类似，我们假设将在针对HPV16E1结合位点的咪唑-吡咯聚酰胺文库中发现几种具有抗病毒活性的化合物。这项第一阶段的研究将建立一个支持HPV16上体维持的角质形成细胞培养系统。聚酰胺将被合成，以靶向 HPV16复制起点(ON)，并测试它们抑制外体DNA水平的能力。为实现这些目标，确定了以下具体目标： 目的1：病毒起源或复制(ORI)将通过合成一系列识别与HPV16相关的规范的E1结合位点的特定序列的聚酰胺来靶向。 目的：在细胞培养中检测聚酰胺对HPV16复制和维持的影响。 目的：HPV16DNA水平下降的阳性结果将得到进一步的鉴定，并为在细胞培养中发挥最大的抗病毒作用创造条件。 这些研究的成功完成为成功的HPV抗病毒治疗提供了希望。 这些研究很重要，因为它们提供了一种替代HPV治疗的疫苗方法。这种方法已经主宰了制药业，而对那些目前感染病毒的人来说却没有任何希望。此外, 这些研究可能确定有价值的研究工具，用于了解HPV生命周期的重要方面。 第二阶段研究将在复杂细胞培养模型(RAFT培养)和HPV相关疾病的动物模型中测试活性化合物的有效性。此外，生物利用度研究将侧重于开发最佳局部治疗配方和测量局部治疗后的全身暴露。这些研究将被要求用于指导毒理学研究。 在人体试验之前。最终，该项目寻求在临床环境中使用针对最流行的致癌病毒(HPV16)的聚酰胺作为局部制剂。

This project seeks imidazole-pyrrole polyamides to be used as topical antiviral agents for the treatment of human papillomavirus (HPV). HPV, especially in its sexually-transmitted forms, presents a major public health problem throughout the world. Current treatments, which include surgery, freezing, noxious agents, and =biological response modifiers," are primitive or non-specific. One reason for the lack of treatment alternatives is the small size of the viral genome, and the corresponding lack of antivirai targets. For this reason, the alternative approach of targeting HPV DNA using sequence-specific DNA binding agents is being taken. The HPV origin of replication (on) contains binding sites for the viral-encoded replication proteins E1 and E2, which are required for viral replication and maintenance. Targeting the or/with sequence-specific DNA binding agents has the potential to block the action of E1 and E2, thereby disrupting processes required for maintenance in cells. Preliminary work with HPV31 has shown significant promise, lmidazole-pyrrole polyamides, designed to target the HPV31 E1 binding site, were found to potently reduce HPV31 episomal DNA levels in cells. We now request supped for the design, synthesis, and testing of compounds that target the commercially important HPV16 genotype. Similar to our preliminary studies with HPV31, we hypothesize that several compounds with antiviral activity will be found among a library of imidazole-pyrrole polyamides targeting the HPV16 E1 binding site. This Phase I study will establish a keratinocyte culture system that supports HPV16 episomal maintenance. Polyamides will be synthesized that target the HPV16 origin of replication (on), and tested for their ability to suppress episomal DNA levels. To meet these objectives, the following specific aims were identified: Aim 1: The viral origin or replication (ori) will be targeted via synthesis of a series of polyamides recognizing the specific sequence of the canonical E1 binding site associated with HPV16. Aim 2: The effects of polyamides on HPV16 replication and maintenance will be tested in cell culture. Aim 3: Positive results showing a decrease in HPV16 DNA levels will be further characterized, and conditions will be established for maximum antiviral effects in cell culture. Successful completion of these studies offers the promise of successful antiviral treatments for HPV. These studies are important because they provide an alternative to the vaccine approaches to HPV therapy. Such approaches have come to dominate the pharmaceutical industry, while offedng no hope to those people currently infected with the virus. In addition, these studies may identifyvaluable research tools for use in understanding important aspects of the HPV life cycle. Phase II studies will test the efficacy of active compounds in complex cell culture models (raft cultures) and animal models of HPV-related disease. In addition, bioavailability studies will focus on developing optimal topical treatment formulations and upon measuring systemic exposure following topical treatments. These studies will be required to guide toxicological studies prior to testing in humans. Ultimately, the project seeks to use polyamides targeting the most prevalent cancer causing virus (HPV16) as topical agents in a clinical setting.