INHIBITORS OF MAST CELLS DEGRANULATION AND TRPM CATION CHANNELS FROM MARINE ORG

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 海洋生物次级代谢产物的结构多样性使其提取物成为生物活性筛选的有效化学库。 拟议研究的总体目标是（a）开发使用高效液相色谱质谱和蒸发光散射检测的高质量预分级海洋天然产物化学库的稳健方法（HPLC-MS-ELSD）和（B）实施这些方法以发现肥大细胞脱粒和TRPM的有效特异性抑制剂。类阳离子通道从海洋生物的提取物。 肥大细胞被认为是炎症性疾病（包括过敏性哮喘和关节炎）的有效效应细胞。 TRPM阳离子通道涉及免疫和神经系统细胞中的一系列细胞功能。 肥大细胞活化的特异性抑制剂或TRPM通道作为治疗或研究工具具有潜在的重要性。 拟议研究的第一个具体目标包括在HPU建立细胞培养实验室，并开发肥大细胞活化和TRPM通道功能的测定方法，可用于筛选海洋生物衍生的化学库。 同时，将开发通过HPLC-MS-ELSD将微生物提取物预分级分离至96孔板格式、复制生物测定板、存档收集的馏分以及分析MS和生物测定数据的方法。 从培养和收集的海洋蓝细菌、藻类和无脊椎动物中获取分类学上不同的样本并将其记录在案的工作将继续进行。 优先级和去复制策略将利用MS配置文件和生物活性，数据库搜索和核磁共振（NMR）光谱之间的相关性。 将通过生物测定和MS定向分级分离高度有希望的提取物中的活性成分，并主要通过NMR和MS进行结构表征。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The structural diversity of secondary metabolites of marine organisms makes their extracts effective chemical libraries for bioassay screening to identify biologically active small molecules. The overall goal of the proposed research is (a) to develop robust methods for the production of high-quality prefractionated marine natural product chemical libraries using high performance liquid chromatography with mass spectrometry and evaporate light scattering detection (HPLC-MS-ELSD) and (b) to implement these methods toward the discovery of potent specific inhibitors of mast cell degranulation and TRPM-class cation channels from extracts of marine organisms. Mast cells are recognized as potent effector cells in inflammatory diseases, including allergic asthma and arthritis. TRPM cation channels are implicated in a range of cellular functions in cells of the immune and nervous systems. Specific inhibitors of mast cell activation, or TRPM channels, are of potential importance as therapeutics or research tools. The first specific aim of the proposed research comprises the establishment of a cell culture laboratory at HPU, and development of assays for mast cell activation and TRPM channel function that can be used to screen marine organism-derived chemical libraries. Concurrently, methods will be developed for the prefractionation of organism extracts by HPLC-MS-ELSD into 96-well plate format, replication of plates for bioassay, archiving of collected fractions, and analysis of MS and bioassay data. Acquisition and documentation of taxonomically diverse samples from cultured and collected marine cyanobacteria, algae, and invertebates will be on-going. Prioritization and dereplication strategies will be employed utilizing correlations between MS profiles and bioactivity, database searches, and nuclear magnetic resonance (NMR) spectroscopy. Active components from highly promising extracts will be isolated by bioassay-and MS-directed fractionation and structually characterized primarily by NMR and MS.