Plasmodium Protein Kinase Focused Antimalarials Discovery

疟疾仍然困扰着大约一半的世界人口，造成40多万人死亡，其中大多数是儿童。是 令人震惊的是，由于疟疾的广泛传播，疟疾治疗的选择越来越有限， 耐药性此外，大多数抗疟药仅作用于红细胞期， 疟疾的预防和复发是次优的。为了解决疟疾治疗的脆弱性，我们建议 发现和优化下一代抗疟药作用于未充分研究的疟原虫激酶。我们将 利用与药物开发相关的各种化学支架和专有技术的丰富知识 和基于激酶通道的疗法对各种人类疾病的耐受性。我们将重点关注II型激酶 抑制剂已经被设计为通过使用ATP结合位点以及 由处于非活性构象的活化环产生的相邻疏水区域。据我们所 尽管目前还不清楚II型抑制剂作为抗疟剂的用途，但仍有待开发。本提案的依据是 在我们有希望的初步筛选，已确定II型化合物与治疗和预防 活动这个项目的目标是测试我们可以设计有效的第二类化学品的假设 支架作用于寄生虫发育的多个阶段。为达致这个目标，我们建议推行 （1）设计和合成优化的选择性抗疟疾II型化合物用于体内应用; 将优化抗疟原虫命中，以提高效力，选择性和药理学特性， 迭代类似物设计、合成和生物和生化活性评价的阶段。（二） 确定先导化合物在小鼠和哺乳动物中的杀灭率、耐药性特征、阶段敏感性和功效， 人源化小鼠模型。此外，先导化合物抑制肝脏阶段感染和 评估配子母细胞成熟。3)我们将通过体外实验确定抗疟化学型的靶点， 耐药性的进化，然后进行全基因组测序。我们将使用条件击倒和复制 用于靶标验证的抗性决定因素的数量变化。我们将进行磷酸化蛋白质组学 分析以揭示所研究的II型抑制剂调节的信号通路。这项研究 这一努力将通过Debopam实验室之间的多学科合作进行 Chakrabarti（中佛罗里达大学），Nathanael Gray（斯坦福大学），Elizabeth Winzeler （加州圣地亚哥大学），在药物化学，激酶化学生物学， 疟疾细胞生物学、抗感染药物发现、靶标鉴定和验证。成功完成 这些目标将导致发现具有治疗和预防潜力的新型化学先导物。

Malaria still afflicts about half of the world population causing more than 400,000 deaths, mostly children. It is quite alarming that the options for malaria therapy are increasingly becoming limited because of widespread drug resistance. Furthermore, most of the antimalarials act only on the erythrocytic stages, the drugs for prophylaxis and relapse of malaria are suboptimal. To address the fragility of malaria therapy, we propose to discover and optimize next generation antimalarials acting on underexplored Plasmodium kinases. We will leverage wealth of knowledge on diverse chemical scaffolds and know-how related to the drug development and tolerability of kinase inhibitor-based therapies for various human diseases. We will focus on type II kinase inhibitors that have been designed to overcome selectivity issues by using both the ATP-binding site as well as the adjacent hydrophobic region created by the activation loop in the inactive conformation. To the best of our knowledge, type II inhibitors are yet to be explored as antimalarial agents. Premise for this proposal is based on our promising preliminary screening that has identified type II compounds with therapeutic and prophylactic activities. The goal of this project is to test the hypothesis that we can design effective type II chemical scaffolds that act on multiple stages of parasite development. To achieve this goal, we propose to pursue the following: (1) Design and synthesize optimized selective antimalarial type II compounds for in vivo application; antiplasmodial hits will be optimized to improve potency, selectivity, and pharmacological properties through stages of iterative analog design, synthesis, and evaluation of biological and biochemical activities. (2) Determine rate of killing, resistance profile, stage susceptibility, and efficacy of lead compounds in murine and humanized mouse models. Furthermore, the ability of the lead compounds to inhibit liver stage infection and gametocyte maturation will be assessed. 3) We will identify targets of antimalarial chemotypes using in vitro evolution of resistance followed by whole genome sequencing. We will use conditional knockdown and copy number variation of the resistance determinants for target validation. We will conduct phosphoproteomics analysis to uncover signaling pathways modulated by the type II inhibitors under study. The research in this endeavor will be conducted through a multidisciplinary collaboration between the laboratories of Debopam Chakrabarti (University of Central Florida), Nathanael Gray (Stanford University), Elizabeth Winzeler (University of California San Diego) with combined expertise in medicinal chemistry, kinase chemical biology, malaria cell biology, anti-infective discovery, target identification, and validation. Successful completion of these goal will lead to the discovery of novel chemical leads with therapeutic and prophylactic potential.