Target-specific antimalarial compound identification using phenotypic assays

我们有效治疗疟疾的能力受到日益广泛的抗药性的威胁， 一线抗疟药物的数量。因此，用于鉴定新的化学探针和/或 迫切需要以生物学验证的靶标为中心的治疗先导物。在这里，我们建议 综合利用功能遗传学和化学生物学方法， 抗疟疾药物的发现。长期目标是利用疟疾寄生虫的最新进展 遗传学，以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。通过关注这一目标类别，我们设想实现发现化学物质的能力。 这些探针能让我们更好地定义基础生物学，阐明抗疟治疗的新选择， 并最终确定有效的治疗策略， 越来越普遍和广泛。虽然基于靶点的药物发现在概念上很有吸引力， 在确定批准的抗疟药物方面，与表型筛选一样成功。但鉴于 大量的基因组学数据和改进的功能遗传学工具， 用于基于目标的发现的方法可以显著地改善这一点。本研究的目的是 (1)建立一种创新的方法来改善靶向特异性药物的发现，同时（2）寻求确定 潜在的探针/药物样分子，其对几种优先目标候选物具有选择性。虽然 相对来说，发现具有体外活性的小分子对靶点是简单的，优先考虑 真正通过该靶点发挥作用以决定生物学结果的化合物一直具有挑战性。相反地， 表型筛选立即确定给定化合物的生物学功效。然而，识别 这些化合物通过其起生物作用的靶是耗时的并且经常是不成功的。无论是 在案例中，能够协同利用经验和理性（例如基于结构的）方法来获取领先优势 化合物优化受到不利影响。在这个建议中，我们整合了最先进的功能， 基因工具将靶特异性信息编码到表型筛选中，以促进快速识别 与预定目标相互作用的化合物。我们寻求建立一个适用于以下方面的普遍框架： 广泛的靶点在其提出的生物学功能、亚细胞定位、生物化学和 生物物理特性我们设想开发标准化的测定和分析管道， 评价各种复合收藏品。目前的建议将侧重于开发和验证低- 至中等通量测定。这项研究意义重大，因为它同时利用了 在疟疾寄生虫中使用靶特异性和表型筛选的关键优势， 潜在的可扩展的过程，可以有助于整体提高效率， 在临床前转化工作期间，开发了有价值的靶标和先导化合物。

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.