New Methods for Simultaneous Arming and SAR Studies of Natural Products

天然产物同时武装和SAR研究的新方法

• 批准号: 7687367
• 负责人: DANIEL ROMO
• 金额: $ 32.43万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-12 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7687367
• 关键词: Alkenes; Alkynes; Amination; Apoptosis; Biological; Biological Assay; Biological Factors; Cellular Assay; Cellular biology; Classification; Collaborations; Complex; Cross-Linking Reagents; Development; Diabetes Mellitus; Disease; Ensure; Genomics; Goals; Grant; Head; High Pressure Liquid Chromatography; Human; Inflammation; Iodination reaction; Lead; Malignant Neoplasms; Mediating; Metals; Methods; Mining; Molecular; Molecular Bank; Monitor; Neurites; Organism; Pathway interactions; Proteins; Reaction; Reagent; Recording of previous events; Research; Scheme; Screening procedure; Site; Structure; T-Cell Activation; Translations; Upper arm; chemical genetics; cycloaddition; design; drug development; functional group; halogenation; human disease; inhibitor/antagonist; metal complex; new therapeutic target; novel; public health relevance; receptor; repository; small molecule; tool

DESCRIPTION (provided by applicant): Natural products have immense structural diversity and interact with a wide range of proteins in diverse organisms and, due to structural homology, also in humans. This proposal seeks to develop a toolbox of reagents and reactions that will enable simultaneous arming and structure- activity studies (SAR) of natural products (NPs). The SAR studies may identify potential new activities of the NP derivatives and will also ensure that initial biological activity has not been lost. While not a Specific Aim of this grant given the scope of this RFA, the arming aspect leads to NP derivatives that are equipped for subsequent conjugation to various tags via Sharpless-Huisgen cycloadditions, enabling biological studies such as mechanism of action (MOA) studies (including off-targets) and provide useful probes for basic cell biology. The long-term objective is to develop a more systemized approach to mine the rich potential of NPs as activators/inactivators of cellular pathways contributing to chemical genetics. The team includes a synthetic organic chemist (D. Romo), an analytical chemist (G. Vigh), a bioorganic/biosynthetic chemist (C. Watanabe), and a biochemist/molecular biologist (J. Liu, Johns Hopkins). We propose to further develop a set of chemo- and site selective reactions employing bifunctional reagents to obtain NP derivatives that are also useful cellular probes. These studies will contribute to a fundamental understanding of the chemo- and site selectivity of various reactions in the context of complex NPs. We have established collaborations with synthetic, biosynthetic, and isolation chemists and they have provided or agreed to provide ~25 bioactive NPs for the proposed studies. Primary reactions to be studied are Rh(II)-promoted O-H insertion reactions, C-H aminations and alkene aziridinations with metal nitrenoids, and mild aryl halogenations. After an initial site non-selective step, we will employ robust/versatile NP derivative purification methods, re- assay the resulting pure derivatives (SAR studies), and determine the most appropriate site for tag attachment, i.e. find the derivatives that retain the greatest bioactivity. We will also screen for potential novel bioactivities of the NP derivatives in various cellular assays and make quantities available to the Molecular Libraries Small Molecule Repository. The subsequent development of both chemo and site-selective versions of the initial reactions will rely on screening various chiral metal complexes, and robust methods for reaction monitoring (LC-MSn) and semi-preparative HPLC purification, to modulate site-selectivity via a type of "double asymmetric synthesis." PUBLIC HEALTH RELEVANCE: The long-term objective of this project is the development of a more systematized approach to mine the rich potential of natural products as activators and inactivators of cellular pathways. Natural products have a rich history as tools for identification of novel therapeutic targets for human disease and as lead structures for drug development. Results from these studies will be directly relevant to human disease including diabetes, inflammation, and cancer since novel methods are proposed for derivatization of natural products (this proposal) pertinent to these disease. Subsequent SAR studies (this proposal) will be pursued to identify potential new bioactivities and suitable sites for probe attachment enabling subsequent mode of action studies (not in this proposal) relevant to these diseases.

描述（由申请人提供）：天然产品具有巨大的结构多样性，并与各种生物体中的广泛蛋白质相互作用，并且由于结构同源性，也与人类相互作用。该提案旨在开发一个试剂和反应工具箱，该工具箱可以同时进行天然产物（NP）的武装和结构活动研究（SAR）。 SAR研究可能会确定NP衍生物的潜在新活动，还将确保最初的生物活性尚未丢失。鉴于此RFA的范围，虽然这不是该赠款的具体目的，但武装方面导致NP衍生物通过Sharpless-Huisgen Cycloaditions随后配备了与各种标签的配置，从而使生物学研究能够进行生物学研究，例如作用机理（MOA）研究（包括非targets）（包括关闭targets），并为基本的细胞生物提供了有用的探针。长期目标是开发一种更系统的方法来挖掘NP作为有助于化学遗传学的细胞途径的激活剂/灭活剂的富裕潜力。该团队包括一名合成有机化学家（D. Romo），一名分析化学家（G. Vigh），一名生物有机/生物合成化学家（C. watanabe）和生物化学家/分子生物学家（J. Liu，J。Liu，Johns Hopkins）。我们建议进一步开发一套采用双功能试剂的化学和位点选择性反应，以获得也是有用的细胞探针的NP衍生物。这些研究将有助于在复杂NP的背景下对各种反应的化学和位点选择性的基本理解。我们已经与合成，生物合成和隔离化学家建立了合作，他们提供或同意为拟议的研究提供约25种生物活性NP。要研究的主要反应是RH（II）促进的O-H插入反应，C-H氨基酶和金属硝酸盐的烯丙基氮杂性以及轻度的芳基卤素化。在最初的站点非选择性步骤之后，我们将采用强大/多功能的NP衍生纯化方法，重新测定所得的纯衍生物（SAR研究），并确定标签附件最合适的位点，即找到保留最大生物活性的衍生物。我们还将在各种细胞测定中筛选NP衍生物的潜在新生物活性性，并使分子库可用的小分子存储库可用。最初反应的化学疗法和现场选择版的随后发展将依赖于筛选各种手性金属复合物，以及用于反应监测（LC-MSN）和半程预留性HPLC纯化的可靠方法，以通过“双重非对称合成”类型来调节现场选择性。 公共卫生相关性：该项目的长期目标是开发一种更加系统化的方法来挖掘天然产品作为激活剂和蜂窝途径灭活剂的丰富潜力。天然产品具有丰富的历史，可以作为鉴定人类疾病的新型治疗靶标的工具和药物开发的铅结构。这些研究的结果将与包括糖尿病，炎症和癌症在内的人类疾病直接相关，因为提出了与这些疾病有关的天然产物（此提案）的新方法。随后的SAR研究（此提案）将被提出，以确定潜在的新生物活性和适当的探测器附着地点，从而实现与这些疾病有关的随后的行动模式研究（不在此提案中）。