Antiviral Compounds that Target HPV18 DNA

描述(由申请人提供)：该项目将确定咪唑-吡咯聚酰胺作为治疗人乳头状瘤病毒(HPV)的局部抗病毒药物。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。我们推测，与我们对HPV16和HPV31的初步研究类似，在针对HPV18E1结合部位的咪唑-吡咯聚酰胺文库中将发现几种具有抗病毒活性的化合物。这一阶段的研究将建立一个角质形成细胞培养系统，以支持HPV18上体的维持。将合成针对HPV18复制起点(ORI)的聚酰胺，并测试它们抑制外体DNA水平的能力。然后，活性化合物将在旨在确认特异性和建立基线选择性指数的测试中进行测试。为实现这些目标，将追求下列具体目标：目标1：化学。HPV18的病毒复制起点(ORI)将通过合成一系列聚酰胺来靶向，这些聚酰胺旨在识别HPV18E1结合位点的特定序列。目的：在细胞培养中检测聚酰胺对HPV18复制和维持的影响。目的3：将进一步鉴定产生HPV18DNA水平降低的化合物，建立在细胞培养中发挥最大抗病毒作用的条件，并进行初步的体外毒性评估。这些研究的成功完成为HPV的特定抗病毒治疗提供了希望。预计降低病毒DNA载量的化合物本身或与体内宿主免疫系统一起，最终足以清除感染HPV的细胞。这些研究很重要，因为它们提供了一种替代HPV疫苗的方法，而HPV疫苗方法已经主导了制药业，而对那些目前或即将感染病毒的人来说没有任何希望。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。未来的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。这些研究将被要求在人体试验之前指导毒理学研究。最终，该项目寻求在临床环境中使用针对最普遍的致癌病毒之一(HPV18)的聚酰胺作为局部制剂。 人乳头瘤病毒是导致宫颈癌的主要原因，宫颈癌是仅次于乳腺癌的女性杀手。目前还没有针对HPV的特效抗病毒治疗。这里描述的研究旨在确定将被用作S局部抗病毒治疗的药物，HPV18型是最重要的致癌病毒之一。

DESCRIPTION (provided by applicant): This project will identify imidazole-pyrrole polyamides to be used as topical antiviral agents for the treatment of human papillomavirus (HPV). Current treatments, which include surgery, freezing, noxious agents, and "biological response modifiers" are primitive or non-specific. We hypothesize that several compounds with antiviral activity will be found among a library of imidazole-pyrrole polyamides targeting the HPV18 E1 binding site similar to our preliminary studies with HPV16 and HPV31. This Phase 1 study will establish a keratinocyte culture system that supports HPV18 episomal maintenance. Polyamides will be synthesized that target the HPV18 origin of replication (ori), and tested for their ability to suppress episomal DNA levels. Active compounds will then be tested in assays designed to confirm specificity and to establish baseline selectivity indices. To meet these objectives, the following specific aims will be pursued: Aim 1: Chemistry. The viral origin of replication (ori) for HPV18 will be targeted by synthesis of a series of polyamides designed to recognize the specific sequence of the HPV18 E1 binding site. Aim 2: The effects of polyamides on HPV18 replication and maintenance will be tested in cell culture. Aim 3: Compounds that produce a decrease in HPV18 DNA levels will be further characterized, conditions will be established for maximum antiviral effects in cell culture, and initial in vitro toxicity assessments will be conducted. Successful completion of these studies offers the promise of specific antiviral treatments for HPV. It is anticipated that compounds reducing viral DNA load will by themselves, or in conjunction with the host immune system in vivo, be ultimately sufficient to clear cells infected with HPV. These studies are important because they provide an alternative to the vaccine approaches to HPV which have come to dominate the pharmaceutical industry, while offering no hope to those people currently or soon to be infected with the virus. In addition, these studies may identify valuable research tools for use in understanding important aspects of the HPV life cycle. Future Phase 2 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 one of the most prevalent cancer causing viruses (HPV18) as topical agents in a clinical setting. Human papillomavirus is the leading cause of cervical cancer, which is second only to breast cancer as a killer of women. There are currently no specific antiviral treatments for HPV. The studies described here are designed to identify agents that will be used as topical antiviral treatment s for HPV 18, one of the most important cancer-causing viruses.