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(申请人提供)：古唑类化合物是一类最近分离的天然产物，通过选择性抑制空泡型ATPase(V-ATPase)对许多人类癌细胞株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。这项拟议研究的长期目标是在彻底了解古拉唑胺抑制活性的基础上创造一类新的小分子V-ATPase抑制剂疗法。本应用的目的是开发一种简化的古唑醇衍生的V-ATPase抑制剂的合成，并使用方便的植物性V-ATPase分析方法进行测定。这是由一个中心假设推动的，即比较系统修饰的古拉西林衍生物的V-ATPase抑制活性将为研究古拉西林的药效团提供关键的见解。这项研究的基本原理是，一旦知道了古拉唑类化合物是如何抑制V-ATPase功能的，就有可能合理地设计具有增强药理学特性的小分子V-ATPase抑制剂，从而为各种疾病的治疗开辟新的途径。在强大的初步数据的指导下，这一假说将通过追求两个具体目标来检验：1)开发简化的青蒿素类似物的合成；2)开发两种以植物为基础的V-ATPase分析。在第一个目标下，以TMS-烯丙化/Peterson消除序列为特征的立体选择性取代三烯合成的合成策略将被用于制备一系列修饰的古唑烷B衍生物，该合成策略已经产生了整个古唑烷碳骨架。然后，将使用在第二个目标下开发的技术来测试高级合成中间体和完成的化合物的V-ATPase抑制活性。具体地说，GFP-拟南芥将被用来鉴定活性化合物，然后可以用拟南芥的根生长试验以剂量依赖的方式进行评估。这项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古唑类化合物的活性，从而背离了V-ATPase研究的传统方法。这一贡献意义重大，因为它旨在阐明V-ATPase结合和抑制的一种新模式。除了提供对V-ATPase结构和功能的进一步了解外，这些结果还有可能导致V-ATPase抑制剂药物的开发，以治疗包括骨质疏松症和癌症在内的各种严重疾病。

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