New Methodologies for Accelerating Natural Product Discovery

描述(申请人提供)：本申请是对天然产物化学新方法申请(RFA-RM-09-005)的回应。我们提出了一项运动，以建立一个新的天然产物发现计划，其基础是a)以洞穴微生物形式存在的新的生物源和b)基于离子迁移质谱仪(IM-MS)的新的分析方法。这两个方面将为天然产物的发现开辟新的生物多样性宝库，并为加速发现粗提物中的化合物并减少其分离所涉及的劳动力提供新的方法。节省的人力和时间来自于能够根据IM-MS测量的性质对代谢物进行分离和结构鉴定的优先顺序。目的1.建立离子迁移质谱仪(IM-MS)分析天然产物的方法。IM-MS是一种二维质谱，它同时绘制了质量/电荷比和气相碰撞截面积，这可能与分子的构象和功能直接相关。我们将评估质量碰撞面积比与自然产品类别的相关性，并将这些相关性与计算预测值进行比较。这些数据将把化合物的结构、类别和构象映射到IM-MS值。目的2.开发洞穴微生物作为天然产物多样性的新来源。可培养的地下粘细菌和放线菌的分离、低温保存和基因分类鉴定将为天然产物的发现提供新的储藏库，并为后续目标提供提取材料来源。目的3.发展IM-MS作为一种在提取物中鉴定候选物质的方法。将通过IM-MS对分类上独特的生物体的子集进行次生代谢潜力评估。具有区别化合物类别、官能度或独特构象的IM-MS性质的化合物将成为分离/结构阐明的目标。这些目标是平行的，但却是协同的。从复杂来源中识别次生代谢物的新方法对于加速发现至关重要。不断需要新的生物多样性来源，以扩大获取生物合成分子多样性的途径。洞穴微生物将提供开发IM-MS作为鉴定萃取物中次级代谢物的工具所需的萃取物的生物来源。通过建议的方法发现的天然产物将提供给美国国立卫生研究院分子图书馆计划(MLP)，包括分子图书馆探针生产中心网络(MLPCN)、分子图书馆小分子仓库(MLSMR)和PubChem。 与公共健康相关：天然产品是药物的极好来源，但它们很难被发现，因为它们必须被确定为生物来源复杂提取物的次要成分。我们建议发展基于分子结构和质量的二维分离，以指导从复杂生物来源中鉴定天然产物。此外，洞穴来源的微生物，一种以前未被开发的生物多样性来源，将被调查以发现天然产品。

DESCRIPTION (provided by applicant): This application is in response to a Request for Applications for New Methodologies in Natural Product Chemistry (RFA-RM-09-005). We propose a campaign to establish a new program of natural product discovery based on a) a new biological source in the form of cave derived microorganisms and b) a new analytical method based on ion mobility mass spectrometry (IM-MS). These two aspects will open up a new reservoir of biodiversity for the discovery of natural products and provide a new method to accelerate the discovery of compounds in crude extracts and reduce the labor involved in their isolation. The labor and time savings derive from the ability to prioritize metabolites for isolation and structure elucidation based on their IM-MS measured properties. Aim 1. Develop natural product analysis using ion mobility mass spectroscopy (IM-MS). IM-MS is a form of 2-dimensional mass spectrometry that plots both mass/charge ratio and gas-phase collision cross- sectional area, which may be directly related to molecular conformation and functionality. We will evaluate the correlation of mass-collision area ratio to natural product class and compare these correlations to computationally predicted values. These data will map compound structure, class and conformation to IM-MS values. Aim 2. Develop cave microorganisms as a new source of natural product diversity. The isolation, cryogenic preservation and genotaxonomic characterization of culturable hypogean myxobacteria and actinobacteria will provide a new reservoir for natural product discovery and a source of extract material for the subsequent aims. Aim 3. Develop IM-MS as a method to identify candidates in extracts. A subset of taxonomically unique organisms will be evaluated for secondary metabolic potential by IM-MS. Compounds with IM-MS properties that distinguish compound class, functionality or unique conformation will be targeted for isolation/structure elucidation. The aims are parallel, but synergistic. New methods for identifying secondary metabolites from complex sources are essential for accelerating discovery. New sources of biodiversity are continuously needed to expand access to their biosynthetic molecular diversity. Cave microorganisms will provide a biological source of extracts needed to develop IM-MS as tool for identification of secondary metabolites in extracts. The natural products discovered by proposed methodologies will be available to the NIH Molecular Libraries Program (MLP) including Molecular Libraries Probe Production Centers Network (MLPCN), Molecular Libraries Small- Molecule Repository (MLSMR) and PubChem. PUBLIC HEALTH RELEVANCE: Natural products are an excellent source for drugs, but they are difficult to discover as they must be identified as minor components of complex extracts from biological sources. We propose to develop two-dimensional separations on the basis of molecular structure and mass to guide the identification of natural products from complex biological sources. Furthermore, cave derived microorganisms, a previously untapped source of biodiversity, will be investigated for natural product discovery.