A chemical proteomic strategy to identify novel drug targets in Plasmodium falciparum and corresponding lead compounds for the development of new antimalarials

疟疾仍然是世界上发病率和死亡率的主要原因之一，每年造成100多万人死亡。鉴于寄生虫对常用药物的广泛耐药性，有必要确定新的药物靶点和开发新的药效团。在目前的联合提案中，我们将利用一种名为基于活性的蛋白质谱(ABPP)的新的化学蛋白质组学策略来识别恶性疟原虫中对生存、生长或致病至关重要的新靶点，以及它们相应的可用于抗疟疾的抑制剂。我们将筛选抑制疟原虫生长的类似天然产物的激发化合物的小的聚焦文库，然后利用ABPP作为化学蛋白质组学方法来鉴定恶性疟原虫及其相应的基本药物靶点。通过这些分析确定的选定的生物相关目标将被克隆并表达重组蛋白。作为初步工作，我们已经确定了一些抑制寄生虫生长的β-内酯化合物，并确定了它们在寄生虫中的潜在蛋白酶靶标。将使用重组蛋白和靶向化合物进行详细的体外相互作用研究和酶活性抑制研究。这些化合物的衍生物将被设计用来识别具有更高亲和力结合/活性抑制和更高抑制寄生虫生长效率的分子。选定的化合物将在小鼠疟疾模型中进一步评估它们抑制寄生虫生长的能力。

Malaria remains one of the leading causes of morbidity and mortality in the world resulting in over a million deaths every year. In view of the widespread development of drug resistance in the parasite against commonly used drugs, there is a necessity to identify new drug targets and develop new pharmacaphores. In the present joint proposal we will utilize a novel chemical proteomic strategy termed activity based protein profiling (ABPP) to identify new targets in P. falciparum that are either essential for survival, growth or pathogenesis together with their corresponding inhibitors that can be developed as anti-malarial. We will screen small focused libraries of drug like natural product inspired compounds that inhibit the parasite growth and then utilize ABPP as chemical-proteomic approach to identify and the corresponding essential drug targets in P. falciparum. Selected biologically relevant targets identified by these analyses will be cloned and recombinant proteins will be expressed. As preliminary work we have already identified some β-lactone compounds that inhibit parasite growth and also identified their potential protease targets in the parasite. Detailed in vitro interaction studies and enzyme activity inhibition studies will be carried out using the recombinant proteins and targeting compounds. Derivatives of these compounds will be designed to identify molecules of higher affinity binding/activity inhibition and higher efficacy to inhibit parasite growth. Selected compounds will be further assessed for their ability to inhibit growth of parasites in a mouse-malaria model.