Antiviral Compounds for Human Papillomavirus 16

该项目寻求咪唑-吡咯聚酰胺用作治疗人乳头瘤病毒（HPV）的局部抗病毒剂。HPV，特别是其性传播形式，在世界各地呈现出一个主要的公共卫生问题。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的尺寸小，并且相应地缺乏抗病毒靶点。出于这个原因，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（上）含有病毒编码的复制蛋白E1和E2的结合位点，这是病毒复制和维持所必需的。靶向序列特异性DNA结合剂或/和序列特异性DNA结合剂具有阻断E1和E2的作用的潜力，从而破坏细胞中维持所需的过程。对HPV 31的初步研究显示出了重要的前景，咪唑-吡咯聚酰胺，旨在靶向HPV 31 E1结合位点， 有效地降低细胞中HPV 31附加型DNA水平。我们现在要求提供设计、合成和测试针对商业上重要的HPV 16基因型的化合物的支持。 与我们对HPV 31的初步研究类似，我们假设在靶向HPV 16 E1结合位点的咪唑-吡咯聚酰胺文库中会发现几种具有抗病毒活性的化合物。本I期研究将建立支持HPV 16游离型维持的角质形成细胞培养系统。聚酰胺将被合成，目标是 HPV 16复制起点（on），并测试它们抑制附加型DNA水平的能力。为实现这些目标，确定了以下具体目标： 目标1：通过合成一系列识别与HPV 16相关的典型E1结合位点的特异性序列的聚酰胺，将靶向病毒起点或复制点（ori）。 目的2：将在细胞培养中测试聚酰胺对HPV 16复制和维持的影响。 目标3：将进一步表征显示HPV 16 DNA水平降低的阳性结果，并建立细胞培养物中最大抗病毒作用的条件。 这些研究的成功完成为HPV的成功抗病毒治疗提供了希望。 这些研究很重要，因为它们为HPV治疗提供了疫苗方法的替代方案。这种方法已经主导了制药业，而对目前感染病毒的人来说却没有希望。此外，本发明还提供了一种方法， 这些研究可能会发现有价值的研究工具，用于了解HPV生命周期的重要方面。 II期研究将测试活性化合物在复杂细胞培养模型（筏培养物）和HPV相关疾病的动物模型中的功效。此外，生物利用度研究将侧重于开发最佳的局部治疗制剂，并测量局部治疗后的全身暴露量。这些研究将被要求指导毒理学研究 在人类测试之前。最终，该项目旨在使用聚酰胺靶向最流行的致癌病毒（HPV 16）作为临床环境中的局部药物。

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.