Anti-infective Natural Products from Deep Sea Vent Organisms

描述（由申请人提供）：在许多人类病原体出现多药耐药性的全球阴影下，新抗生素的管道正在惊人地耗尽。为了解决这一紧迫的公共卫生问题，必须不断寻求通过新机制起作用的新型结构新颖分子。在过去的25年里，超过50%的新化学实体被批准为药物是基于天然产物先导化合物。因此，化学上不同的天然产物来自不同的，系统发育独特的生物是下一代治疗剂的关键线索。本课题的总体目标是利用微尺度化学和生物技术对新发现的深海喷口生物进行评价，以获得具有医学相关抗感染特性的前所未有的小分子天然产物，并将实现以下具体目标：1)获得和化学提取野外采集和实验室培养的深海喷口生物。2)利用液相色谱-质谱（LC-MS）和自动微流毛细管核磁共振（NMR）谱技术对vent样品的HPLC组分库进行化学分析。3)对vent样品库进行体外抗菌活性生物学分析，并在体外细菌耐药试验和体内快速、容易进行的斑马鱼感染模型中测试纯化合物先导物。深海潜水车收集的深海喷口无脊椎动物和微生物垫将进行小规模的提取、过滤和高效液相色谱分离，以组装一个高效液相色谱馏分的深海喷口样本库。此外，深喷口细菌的实验室培养受到相同的协议将有助于样本库。分析数据包括紫外吸收，质谱和核磁共振光谱数据将为HPLC馏分库获得，以便快速识别已知或潜在的新型化合物。通过低重复高效液相色谱运行产生的额外数量的这些高效液相色谱组分将在微滴液体培养试验中对临床相关的革兰氏阳性和革兰氏阴性细菌进行测试。在这些检测中具有活性的高效液相色谱纯化化合物将提交给对多重耐药细菌病原体的进一步评估。此外，抗菌纯化合物引线将在斑马鱼胚胎分枝杆菌和葡萄球菌感染的体内模型中进行测试。

DESCRIPTION (provided by applicant): In the global shadow of emerging multi-drug resistance in many human pathogens, the pipeline of new antibiotics is alarmingly depleted. To address this pressing public health concern, new types of structurally novel molecules that act via new mechanisms must constantly be sought. In the last 25 years, over 50% of all new chemical entities approved as drugs were based on a natural product lead compound. Thus, chemically diverse natural products from diverse, phylogenetically unique organisms are critical leads for the next generation of therapeutic agents. The overarching goal of this proposal is to use microscale chemical and biological techniques to evaluate recently discovered deep-sea vent organisms as a source of unprecedented small molecule natural products with medicinally-relevant anti-infective properties, and will be achieved by the following specific aims: 1) To obtain and chemically extract field-collected and laboratory-cultured deep vent organisms. 2) To chemically profile a library of vent sample HPLC fractions by liquid chromatograph-mass spectrometry (LC-MS) and automated microflow capillary nuclear magnetic resonance (NMR) spectroscopy. 3) To biologically profile the vent sample library for antimicrobial activity in vitro and to test pure compound leads in vitro in bacterial resistance assays and in vivo in rapid, easily performed zebrafish infection models. Deep vent invertebrates and microbial mats collected by deep submergence vehicles will be subjected to small-scale extraction, filtration and HPLC fractionation to assemble a vent sample library of HPLC fractions. Additionally, laboratory cultures of deep vent bacteria subjected to the same protocols will contribute to the sample library. Analytical data including UV absorption, mass spectrometric and NMR spectroscopic data will be obtained for the HPLC fraction library to enable rapid identification of known or potentially new types of chemical compounds. Additional quantities of these HPLC fractions generated by low replicate HPLC runs will be tested in microtiter liquid culture assays against clinically relevant Gram-positive and Gram-negative bacteria. Purified compounds from HPLC fractions with activity in these assays will be submitted for further evaluation against multi-drug resistant bacterial pathogens. In addition, antibacterial pure compound leads will be tested in in vivo models of mycobacterial and staphylococcal infections of embryonic zebrafish. PUBLIC HEALTH RELEVANCE: The treatment of infectious diseases with antibiotics is plagued by the rapid and inevitable development of drug-resistance in the disease causing microorganisms, and requires the constant discovery of new drug leads that produce their antibiotic effect in new ways. This research focuses on the evaluation of unusual, recently discovered deep-sea hydrothermal vent organisms for their production of novel types of natural products for anti-infective drug development. Natural products are the source of the majority of approved antibiotic drugs, and are also a proven source of new types of chemical compounds.