Small Molecule Inhibitors of EBV Latency

EBV潜伏期的小分子抑制剂

• 批准号: 6656740
• 负责人: JEFFERY T SAMPLE
• 金额: $ 19.9万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6656740
• 关键词: DNA binding protein; DNA replication; Epstein Barr virus; antiinfective agents; binding sites; combinatorial chemistry; gel mobility shift assay; gene expression; genetic transcription; high throughput technology; inhibitor /antagonist; latent virus infection; microorganism disease chemotherapy; oncogenic virus; oncoproteins; polymerase chain reaction; technology /technique development; virus related neoplasm /cancer

DESCRIPTION (provided by applicant): The objective of the proposed research is to evaluate the antiviral potential of synthetic small molecule compounds known as polyamides for their ability to specifically inhibit Epstein-Barr virus (EBV) latency-gene expression and functions that contribute to EBV-associated lymphomas and lymphoproliferative disease in immune compromised individuals. Polyamides bind within the minor groove of a DNA double helix in a sequence-specific manner with affinities comparable to many sequence-specific DNA-binding proteins. Consequently, these compounds have the ability to compete with proteins (e.g., transcription factors) that target the same or an overlapping binding site within dsDNA, thus disrupting their function. This and the generally favorable pharmacological properties of polyamides make them attractive as antiviral drugs to inhibit the expression or DNA-binding functions of viral proteins critical for pathogenesis. Such agents targeting latent EBV infection are particularly attractive, since current anti-herpesvirus drugs are designed to inhibit virus replication and are ineffective against the latent or transforming form of EBV infection. Three specific aims are proposed. Under Aim 1 we will evaluate the in vivo inhibitory potential of hairpin polyamides designed to target EBV latency gene promoters and the viral origin of DNA replication, oriP. Specifically, we will seek to block transcription of the EBV genes encoding the essential genome maintenance protein EBNA-1 and the oncoprotein LMP-1 by inhibition of the binding of transcription factors to critical cis-regulatory elements of the EBNA- 1 and LMP- 1 promoters. Additionally, we will test whether polyamides designed to inhibit binding of EBNA- 1 to oriP are capable of enforcing loss of the EBV genome from EBV-positive lymphoma cells. Under Aim 2 we propose to develop DNA-binding and whole-cell reporter based assays that are compatible with high throughput screening of chemical libraries so that the most effective inhibitors of EBNA-1 binding to oriP and of EBNA-1 and LMP-1 expression can be identified. Under Aim 3, we will take a combinatorial approach to identify polyamide inhibitors of EBNA-1 DNA-binding and of EBNA-1 and LMP-1 expression. Specifically, we propose to generate a combinatorial library of polyamide-based compounds that will be screened, using the bioassays developed under Aim 2, to identify lead compounds with anti-EBV properties. By taking the complementing approaches of rational and combinatorial drug design in Aims 1 and 3, respectively, we hope to effectively evaluate the anti-EBV potential of these compounds, and identify lead compounds with potential therapeutic value.

描述（由申请人提供）： 拟议的研究的目的是评估合成小分子化合物的抗病毒潜力，称为聚酰胺，其特异性抑制EPSTEIN-BARR病毒（EBV）潜伏期的表达和功能有助于EBV相关的淋巴瘤和免疫疾病中的淋巴细胞增生性疾病。聚酰胺以序列特异性的方式结合DNA双螺旋的小凹槽，其亲和力与许多序列特异性DNA结合蛋白相当。因此，这些化合物具有靶向dsDNA中相同或重叠结合位点的蛋白质（例如转录因子）竞争的能力，从而破坏了它们的功能。这种和普遍有利的聚酰胺药理特性使它们作为抗病毒药物具有吸引力，可抑制病毒蛋白的表达或DNA结合功能对发病机理至关重要。这种靶向潜在EBV感染的药物特别有吸引力，因为当前的抗hepevirus药物旨在抑制病毒复制，并且对EBV感染的潜在或转化形式无效。提出了三个具体目标。在AIM 1下，我们将评估旨在靶向EBV潜伏基因启动子和DNA复制的病毒起源的发夹聚酰胺的体内抑制潜力。具体而言，我们将寻求通过抑制转录因子与EBNA-1和LMP-1启动子的关键顺式调节元件的结合来阻止编码基本基因组维持蛋白EBNA-1和癌蛋白LMP-1的EBV基因的转录。此外，我们将测试旨在抑制EBNA-1与Orip结合的聚酰胺是否能够从EBV阳性淋巴瘤细胞中执行EBV基因组丧失。在AIM 2下，我们建议开发与化学文库高吞吐量筛选兼容的DNA结合和全细胞报告基因测定法，以便可以鉴定出EBNA-1最有效的抑制剂，以及可以鉴定出EBNA-1和LMP-1表达的抑制剂。在AIM 3下，我们将采用一种组合方法来鉴定EBNA-1 DNA结合以及EBNA-1和LMP-1表达的聚酰胺抑制剂。具体而言，我们建议生成一个基于聚酰胺化合物的组合库，该化合物将使用AIM 2下开发的生物测定筛选，以识别具有抗EBV特性的铅化合物。通过分别采用AIM 1和3中理性和组合药物设计的补充方法，我们希望有效地评估这些化合物的抗EBV潜力，并鉴定具有潜在治疗价值的铅化合物。