Probing the Archazolid Pharmacophore Using Plant-Based V-ATPase Assays

使用基于植物的 V-ATP 酶测定法探测 Archazolid 药效团

• 批准号: 8290763
• 负责人: Gregory W O'Neil
• 金额: $ 29.03万
• 依托单位: WESTERN WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8290763
• 关键词: ATP phosphohydrolase; Arabidopsis; Binding; Biological Assay; Biological Factors; Cancer cell line; Carbon; Complex; Data; Development; Disease; Dose; Family; Fostering; Goals; Growth; Health; Human; Knowledge; Lead; Malignant Neoplasms; Mission; Osteoporosis; Outcome; Pharmaceutical Preparations; Plant Roots; Plants; Property; Public Health; Renal tubular acidosis; Research; Series; Side; Structure; Techniques; Testing; Therapeutic; Virus Diseases; Work; analog; base; burden of illness; design; high throughput screening; human disease; improved; inhibitor/antagonist; innovation; insight; novel; novel strategies; pharmacophore; prevent; scaffold; small molecule; tool; tool development; vacuolar H+-ATPase

DESCRIPTION (provided by applicant): The archazolids are a family of recently isolated natural products that display powerful growth inhibitory activity against a number of human cancer cell lines based on selective vacuolar-type ATPase (V-ATPase) inhibition, however the mechanism by which these compound achieve their activity is not completely understood. A long-term goal of the proposed research is to create a new class of small-molecule V-ATPase inhibitor therapeutics based on a thorough understanding of archazolid inhibitory activity. The objective of this application is to develop a synthesis of simplified archazolid-derived V-ATPase inhibitors and assay using convenient plant-based V-ATPase assays. This is driven by a central hypothesis that the comparison of the V-ATPase inhibitory activity of systematically modified archazolid-derivatives will provide crucial insights into the archazolid pharmacophore. The rationale for the proposed research is that once it is known how the archazolids inhibit V-ATPase function, it will become possible to rationally design small molecule V-ATPase inhibitors with enhanced pharmacological properties resulting in new approaches for the treatment of various diseases. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Develop a synthesis of simplified archazolid analogues; and 2) Develop two plant-based V-ATPase assays. Under the first aim, a synthetic strategy featuring a TMS-allylation/Peterson elimination sequence for stereoselective substituted triene synthesis that has already produced the entirety of the archazolid carbon framework will be used to prepare a series of modified archazolid B derivatives. The V-ATPase inhibitory activity of advanced synthetic intermediates and completed compounds will then be tested using the techniques developed under the second aim. Specifically, GFP-Arabidopsis will be used to identify active compounds that can then be evaluated in a dose-dependent manner using an Arabidopsis root-growth assay. The proposed research is innovative because it represents a departure from the traditional plecomacrolide approach to V-ATPase research by providing research tools to better elucidate archazolid activity. This contribution is significant because it aims to elucidate a nove mode of V-ATPase binding and inhibition. In addition to providing further insights into V-ATPase structure and function, the results have the potential to lead to new approaches for the development of V-ATPase inhibitor drugs to treat various severe diseases including osteoporosis and cancer.

描述（由申请人提供）：archazolid 是最近分离的天然产物家族，基于选择性液泡型 ATP 酶（V-ATP 酶）抑制，对多种人类癌细胞系显示出强大的生长抑制活性，但这些化合物实现其活性的机制尚不完全清楚。该研究的长期目标是基于对 archazolid 抑制活性的透彻了解，创建一类新型小分子 V-ATP 酶抑制剂疗法。本申请的目的是开发一种简化的 archazolid 衍生的 V-ATP 酶抑制剂的合成方法，并使用方便的基于植物的 V-ATP 酶测定进行测定。这是由一个中心假设驱动的，即系统修饰的 archazolid 衍生物的 V-ATP 酶抑制活性的比较将为 archazolid 药效团提供重要的见解。这项研究的基本原理是，一旦了解了archazolids如何抑制V-ATP酶功能，就可以合理设计具有增强药理特性的小分子V-ATP酶抑制剂，从而为治疗各种疾病提供新方法。在强有力的初步数据的指导下，这一假设将通过追求两个具体目标进行检验：1）开发简化的阿唑烷类似物的合成； 2) 开发两种基于植物的 V-ATP 酶测定法。第一个目标是采用TMS-烯丙基化/Peterson消除序列进行立体选择性取代三烯合成的合成策略，该策略已经产生了整个archazolid碳骨架，将用于制备一系列改性archazolid B衍生物。然后将使用第二个目标开发的技术测试高级合成中间体和完整化合物的 V-ATP 酶抑制活性。具体而言，GFP-拟南芥将用于鉴定活性化合物，然后使用拟南芥根生长测定以剂量依赖性方式对其进行评估。拟议的研究具有创新性，因为它通过提供研究工具来更好地阐明 archazolid 活性，代表了 V-ATP 酶研究中传统的 plecomacrolide 方法的背离。这一贡献意义重大，因为它旨在阐明 V-ATP 酶结合和抑制的新模式。除了提供对 V-ATP 酶结构和功能的进一步了解外，这些结果还有可能为开发 V-ATP 酶抑制剂药物以治疗包括骨质疏松症和癌症在内的各种严重疾病提供新方法。

项目成果

Synthesis of a C1-C23 fragment of the archazolids and evidence for V-ATPase but not COX inhibitory activity.

archazolid 的 C1-C23 片段的合成以及 V-ATP 酶而非 COX 抑制活性的证据。

• DOI: 10.1055/s-0036-1588413
• 发表时间: 2017
• 期刊: Synlett : accounts and rapid communications in synthetic organic chemistry
• 作者: O'Neil,GregoryW;Craig,AlexanderM;Williams,JohnR;Young,JeffreyC;Spiegel,PClint
• 通讯作者: Spiegel,PClint

Synthesis of the C1-C17 fragment of the archazolids by complex cis-homodimer cross metathesis.

通过复杂的顺式同二聚体交叉复分解合成 archazolid 的 C1-C17 片段。

• DOI: 10.1016/j.tetlet.2015.05.014
• 发表时间: 2015
• 期刊: Tetrahedron letters
• 影响因子: 1.8
• 作者: Swick,StevenM;Schaefer,SaraL;O'Neil,GregoryW
• 通讯作者: O'Neil,GregoryW

Exploring a Ring-Closing Metathesis Approach to the Archazolid Macrocycle.

探索 Archazolid 大环的闭环复分解方法。

• DOI: 10.1055/s-0034-1379003
• 发表时间: 2014
• 期刊: Synthesis
• 作者: King,BrianneR;Swick,StevenM;Schaefer,SaraL;Welch,JamieR;Hunter,EmilyF;O'Neil,GregoryW
• 通讯作者: O'Neil,GregoryW