DNA Polymerase IIIe, A New Antibiotic Target

DNA 聚合酶 IIIe，新的抗生素靶点

• 批准号: 7038266
• 负责人: George E Wright
• 金额: $ 90.71万
• 依托单位: GLSYNTHESIS, INC.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-11-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7038266
• 关键词: DNA directed DNA polymerase; Enterococcus; Staphylococcus aureus; Streptococcus pneumoniae; Streptococcus pyogenes; X ray crystallography; active sites; antibacterial agents; bacterial proteins; biotechnology; biotherapeutic agent; chemical synthesis; cytotoxicity; drug design /synthesis /production; drug discovery /isolation; drug interactions; drug resistance; drug screening /evaluation; enzyme inhibitors; guanine analog; guinea pigs; high performance liquid chromatography; laboratory mouse; nuclear magnetic resonance spectroscopy; pharmacokinetics

DESCRIPTION (provided by applicant): In phase I we have identified a new class of active-site directed inhibitors of DMA polymerase HIE, a novel replicative enzyme in Gram+ bacteria. The compounds, 7-substituted-N2-(3,4-dichlorobenzyl)guanines "DCBGs", are potent enzyme inhibitors, and selected derivatives have potent and broad activity against clinically relevant Gram+ bacteria. We have identified the first active site directed inhibitors of the related DNA polymerase HIE from Gram- bacteria, i.e. E. coli. In addition, we have established a collaboration to crystallize and solve the structure of a complex between E. fecalis pol IIIE, DNA and one of our inhibitors. Based on the results of phase I, we will pursue designation of a lead antibiotic compound by in vivo testing in systemic and toplical bacterial infection models. We will use both QSAR analysis and structure-based drug design to discover new platform inhibitors of the pol HIE target. The following specific aims will be pursued: 1. scale-up synthesis of lead compound candidates and formulations for evaluation in animal models of bacterial infection; synthesis of 7-substituted analogs with enhanced pol HIE inhibitory and antibacterial activity in vitro. 2. solve the structure of E. fecalis pol IIIE:DNA:inhibitor complexes, and use the coordinates for understanding the basis of inhibition of the enzyme and for further rational, computer-based drug design (collaboration with Dr. Mark Jedrzejas, Children's Hospital of Oakland Research Institute). 3. continue assays of compounds for enzyme inhibition (pol IIIC, pol HIE), antibacterial activity, selectivity, cytotoxicity; incidence of resistance and mechanism(s) of resistance, by cloning and sequencing of targets (by subcontract to Microbiotix Inc.); test candidate drugs against clinical isolates of Gram+ and Gram- bacteria, bactericidal assays, combination studies with marketed antibiotics (by subcontract to UMass Medical School). 4. develop analytical methods for analysis of candidates in animal plasma, drug uptake and distribution by various routes in mice, acute toxicity, in vitro metabolism and stability studies. 5. evaluate candidate drugs for activity in animal models of Gram+ and Gram- infections (if warranted), emphasizing systemic antibiotic-resistant S. aureus, E. fecalis and S. pneumoniae in mice and topical S. aureus infections in guinea pigs.. Potent inhibition of Gram+ pol HIE and pol IIIC may lead to antibiotics with reduced incidence of resistance.

描述（由申请人提供）：在第一阶段，我们确定了一种新的活性位点定向抑制剂DMA聚合酶HIE，这是一种新型的革兰氏+细菌中新型复制酶。这些化合物是7-取代的N2-（3,4-二氯苯基）鸟嘌呤“ DCBGS”，是有效的酶抑制剂，所选的衍生物具有对临床相关的革兰氏+细菌的有效和广泛的活性。我们已经确定了第一个活跃的位点定向的抑制剂，该抑制剂来自革兰氏菌，即大肠杆菌。此外，我们还建立了协作，以结晶和解决粪肠球菌，DNA和我们的抑制剂之一之间的复合物结构。 根据第一阶段的结果，我们将通过体内和全身细菌感染模型中的体内测试来追求铅抗生素化合物的名称。我们将同时使用QSAR分析和基于结构的药物设计来发现POL HIE目标的新平台抑制剂。将追求以下具体目标： 1。在细菌感染动物模型中评估铅复合候选物和制剂的扩大合成；在体外，与增强的pol Hie抑制性和抗菌活性的7个取代类似物合成。 2。解决大肠杆菌的结构：DNA：DNA：抑制剂复合物，并利用坐标来理解抑制酶的基础，并进一步基于计算机的药物设计（与奥克兰研究所儿童医院的Mark Jedrzejas博士合作）。 3。继续测定酶抑制的化合物（POL IIIC，POLHIE），抗菌活性，选择性，细胞毒性；通过克隆和测序靶标（通过分包给Microbobiotix Inc.），电阻和机制的发生率和机制的发生率；测试候选药物针对革兰氏+和革兰氏菌细菌的临床分离株，杀菌测定，与市场抗生素的组合研究（分包给UMass医学院）。 4。开发分析方法，用于分析动物血浆中的候选物，通过小鼠，急性毒性，体外代谢和稳定性研究的各种途径的药物吸收和分布。 5。评估候选药物的革兰氏+和革兰氏阴性动物模型的活性（如果有必要），强调了全身性抗生素的金黄色葡萄球菌，E。fecalis和S. pneumoniae的小鼠以及豚鼠中的金黄色葡萄球菌感染。 对革兰氏+ pol Hie和pol IIIC的有效抑制可能导致抗生素，耐药性降低。