Anti-infective Natural Products from Deep Sea Vent Organisms

描述(申请人提供)：在许多人类病原体出现多重耐药性的全球阴影下，新抗生素的流水线令人担忧地耗尽。为了解决这一紧迫的公共卫生问题，必须不断寻找通过新机制发挥作用的新型结构新颖分子。在过去的25年里，所有被批准为药物的新化学实体中，超过50%是以天然产品铅化合物为基础的。因此，来自不同的、系统发育独特的生物体的化学上不同的天然产品是下一代治疗剂的关键线索。这项提议的总体目标是利用微型化学和生物学技术来评估最近发现的深海喷口生物作为具有药物相关抗感染特性的史无前例的小分子天然产品的来源，并将通过以下具体目标实现：1)获得并以化学方法提取现场收集和实验室培养的深海喷口生物。2)通过液-质联用(LC-MS)和自动微流控毛细管核磁共振(核磁共振)谱对VET样品的高效液相色谱库进行化学图谱分析。3)对Vent样本库的体外抗菌活性进行生物学分析，并在体外细菌耐药性检测和快速、易于操作的斑马鱼感染模型中测试纯化合物先导化合物。深水潜水器采集的深水喷口无脊椎动物和微生物垫将经过小规模提取、过滤和高效液相色谱分级，以组装喷口高效液相色谱组分的样本库。此外，符合相同方案的深喷口细菌的实验室培养将为样本库做出贡献。将获得包括紫外线吸收、质谱学和核磁共振波谱数据在内的分析数据，以便能够快速识别已知或潜在的新类型的化合物。通过低重复的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针对临床相关的革兰氏阳性和革兰氏阴性细菌的微滴度液体培养分析中进行测试。从高效液相色谱级分中提纯的具有这些检测活性的化合物将提交进一步评估，以对抗多重耐药细菌病原体。此外，抗菌纯化合物先导将在斑马鱼胚胎的分枝杆菌和葡萄球菌感染的体内模型中进行测试。 与公共卫生相关：抗生素治疗传染病受到致病微生物耐药性快速和不可避免的发展的困扰，需要不断发现以新方式产生抗生素效果的新药物先导。这项研究的重点是对最近发现的不寻常的深海热液喷口生物进行评估，以生产用于抗感染药物开发的新型天然产品。天然产品是大多数已获批准的抗生素药物的来源，也是经证实的新型化合物的来源。

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.