Target-specific antimalarial compound identification using phenotypic assays

我们有效治疗疟疾的能力受到对有限数量的日益广泛的耐药性的威胁 可供选择的一线抗疟疾药物。因此，用于识别新的化学探针和/或 迫切需要以生物验证靶点为中心的治疗线索。在这里，我们建议 综合使用功能遗传学和化学生物学方法，以实现更有效的目标驱动 抗疟疾药物的发现。长期目标是利用疟疾寄生虫方面的最新进展。 遗传学-鉴定和优先考虑以前未探索的生物靶点和/或治疗途径 探索的努力。通过专注于这一目标类别，我们设想实现发现化学物质的能力 更好地让我们定义基础生物学和阐明抗疟疾治疗的新选择的探测， 并最终确定有效的治疗策略来对抗正在成为 变得越来越普遍和普遍。尽管基于靶点的药物发现在概念上很有吸引力，但它并没有 在鉴定经批准的抗疟疾药物方面与表型筛查一样成功。然而，鉴于 大量的基因组数据和改进的功能遗传学工具现在可用，更有效 基于目标的发现方法可以极大地改善这一点。这项研究的目的是 (1)建立一种创新的方法来改进靶向药物发现，同时(2)寻求确定 潜在的探针/类药物分子，对几个优先目标候选具有选择性。当它 相对直截了当地发现具有体外活性的小分子针对目标，优先 真正通过这个目标来决定生物结果的化合物一直是具有挑战性的。相反， 表型筛选可立即确定特定化合物的生物功效。但是，识别 这些化合物通过其生物作用的目标(S)是耗时的，而且往往不成功。在这两个地方 在案例中，能够协同利用经验和理性(例如，基于结构的)铅方法 复合优化受到了不利影响。在这个提案中，我们集成了最先进的功能 基因工具将靶标特定信息编码到表型屏幕中，以便于快速识别 与预先指定的目标相互作用的化合物。我们试图建立一个通用的框架，适用于 广泛的靶标在其拟议的生物学功能、亚细胞定位、生化和 生物物理特性。我们设想开发标准化的分析和分析管道，以促进 各种复合藏品的评价。目前的提案将侧重于开发和验证Low-Low。 中等量分析。这项拟议的研究意义重大，因为它同时利用 在疟疾寄生虫中使用靶标特异性和表型筛查的关键优势成为快速、稳健和 潜在的可扩展过程，可总体上有助于提高治疗效率 有价值的靶点和先导化合物在临床前的转化工作中被提出。

Our ability to effectively treat malaria is threatened by increasingly widespread resistance to the limited number of frontline antimalarial drugs available. Therefore, new strategies for identifying novel chemical probes and/or therapeutic leads centered around biologically validated targets are critically needed. Here, we propose the integrated use of functional genetics and chemical biology approaches to enable more effective target-driven antimalarial drug discovery. The long-term goal is to take advantage of recent advances in malaria parasite genetics to qualify and prioritize previously unexplored biological targets and/or pathways for therapeutics discovery efforts. By focusing on this target category, we envision achieving the capability to discover chemical probes that better allow us to define fundamental biology and elucidate new options for antimalarial therapy, and ultimately identify effective therapeutic strategies against multidrug resistant parasites that are becoming increasingly prevalent and widespread. While target-based drug discovery is conceptually appealing, it has not been as successful as phenotypic screens in identifying approved antimalarial agents. However, given the substantial amount of genomics data and the improved functional genetics tools now available, more effective approaches for target-based discovery could dramatically improve this. The objectives of this research are to (1) establish an innovative approach for improving target-specific drug discovery while (2) seeking to determine potential probe/drug-like molecules that are selective against several prioritized target candidates. While it relatively is straightforward to discover small molecules with in vitro activity against a target, prioritizing compounds that truly act via that target to dictate the biological outcome has been challenging. Conversely, phenotypic screens immediately establish the biological efficacy of a given compound. However, identifying the target(s) through which these compounds act biologically is time consuming and often unsuccessful. In both cases, the ability to synergistically leverage empirical and rational (e.g. structure-based) approaches for lead compound optimization is adversely impacted. In this proposal, we integrate use of state-of-the-art functional genetic tools to encode target-specific information into phenotypic screens to facilitate rapid identification of compounds interacting with pre-specified targets. We seek to establish a generalized framework applicable to a broad range of targets varying in their proposed biological function, subcellular localization, biochemical and biophysical properties. We envision developing standardized assays and analytical pipelines to facilitate evaluation of various compound collections. The current proposal will focus on developing and validating low- to-moderate throughput assays. The proposed research is significant because it simultaneously leverages key strengths of using target-specific and phenotypic screens in malaria parasites into a rapid, robust and potentially scalable process that can contribute overall to improving the efficiency with which therapeutically valuable targets and lead compounds are advanced during preclinical translational efforts.