Hydroquinone derivatives as novel calcium ATPase inhibitors

描述（由申请人提供）：源自天然产物毒胡萝卜素（TG）的肌浆细胞/内质网钙-ATP酶（SERCA）的常用抑制剂受到归因于其高结构复杂性的限制，这限制了其可用性并使其通过有机合成的定制具有挑战性和繁琐性。由于它们作为研究SERCA在生理过程中的作用的强大研究工具的价值以及它们作为抗前列腺癌的新型药剂的潜力，开发基于TG的SERCA抑制剂的替代品是非常可取的。本提案中概述的研究是基于对苯二酚（HQ）结构的新型SERCA抑制剂的设计和深入表征。尽管它们的尺寸较小且复杂性较低，但基于HQ的抑制剂-虽然容易获得且方便定制-显示出针对SERCA活性的显著高效力。作为实现全面了解SERCA与小抑制分子相互作用的长期目标的重要一步，该特定应用的目的是发现和全面表征HQ基抑制剂，这些抑制剂是未来发展为研究工具或抗癌剂的良好候选物。初步数据支持中心假设，即进一步将HQ开发为新的SERCA抑制剂类别是一项可行的任务，并可能产生新的研究工具和/或抗癌药物。预测具有良好效力的抑制剂将被合成或通过化合物文库筛选获得，然后在生物测定中测量其效力。还将评估将肽系链连接至SERCA抑制剂以传递对前列腺癌细胞的特异性的可行性。作为合成、文库筛选和生物测定的补充，将通过分子动力学（MD）模拟对最有效化合物的选择进行彻底表征，以便在分子水平上提供分子间相互作用和结合位点事件的时间依赖性说明。在一些晶体学工作的支持下，MD模拟还将预测SERCA抑制剂的结合亲和力，以帮助指导新化合物的合成。此外，SERCA抑制剂干扰酶在生命系统中的Ca 2+转运功能的能力将在健康和癌细胞的成像研究中进行评估。拟议的研究是创新的，因为它将探索HQ支架作为基于TG的SERCA抑制剂的有吸引力的替代品。此外，它的重要性源于其产生基本信息的目标，这些信息对于最终将HQ开发成有价值的新研究工具和抗前列腺癌的新药物是必要的。通过让本科研究生参与多学科药物设计项目，计划中的项目符合NIH AREA计划的目标。

DESCRIPTION (provided by applicant): Commonly used inhibitors of the enzyme sarco/endoplasmic reticulum calcium-ATPase (SERCA) derived from the natural product thapsigargin (TG) suffer from limitations attributed to their high structural complexity, which restricts their availability and makes their customization by organic synthesis challenging and cumbersome. Because of their value as powerful research tools for the study of SERCA's role in physiological processes and their potential as novel agents against prostate cancer, the development of alternatives to TG-based SERCA inhibitors is highly desirable. The research outlined in this proposal is the design and the in-depth characterization of novel SERCA inhibitors based on the structure of hydroquinone (HQ). Despite their smaller size and lower complexity, HQ- based inhibitors - while readily available and conveniently customized - display remarkably high potencies against SERCA activity. As an important step towards the long-term goal of obtaining a comprehensive understanding of SERCA's interaction with small inhibitory molecules, the objective of this particular application is the discovery and comprehensive characterization of HQ-based inhibitors that are good candidates for the future development into research tools or anti-cancer agents. Preliminary data support the central hypothesis that the further development of HQs into a new SERCA inhibitor class is a feasible task and will likely yield novel research tools and/or anti-cancer agents. Inhibitors that are predicted to have good potencies will be synthesized or obtained by compound library screens and their potencies will then be measured in bioassays. The feasibility of attaching peptide tethers to SERCA inhibitors to convey specificity for prostate cancer cells will also be evaluated. Complementary to synthesis, library screens, and bioassays, a selection of the most potent compounds will be characterized thoroughly by molecular dynamics (MD) simulations in order to furnish a time-dependent account of the intermolecular interactions and events in the binding site at the molecular level. Supported by some crystallographic work, the MD simulations will also predict binding affinities of inhibitors for SERCA in order to help guide the synthesis of new compounds. Furthermore, the ability of SERCA inhibitors to interfere with the enzyme's Ca2+ transport function in living systems will be assessed in imaging studies with healthy and cancerous cells. The proposed research is innovative because it will explore the HQ scaffold as an attractive alternative to TG-based SERCA inhibitors. Moreover, its significance stems from its goal of generating fundamental information that is necessary to eventually develop HQs into valuable new research tools and into novel agents against prostate cancer. By engaging undergraduate research students in a multi-disciplinary drug design project, the planned project meets the goals of the NIH AREA program.