Apicomplexan Drug Target Discovery

描述（由申请人提供）：顶复门寄生虫包括重要的人类病原体，如疟原虫属，弓形虫和隐孢子虫。由于缺乏疫苗以及不断发展的耐药性和严重的副作用，目前可用的治疗，需要新的药物来治疗这些感染。虽然高通量小分子筛选已经确定了许多抗疟疾的先导化合物，但尚不清楚几乎所有这些化合物如何杀死疟原虫。了解这些机制很重要，原因有两个。首先，将一种化合物开发成可以开给患者的药物需要彻底了解化合物的作用机制。其次，药物通过抑制细胞化合物和途径发挥作用，发现这些化合物和途径揭示了新的药物靶点。由于许多顶复门特有的过程对这些寄生虫的生长至关重要，因此一些抗疟疾先导化合物可能也会影响其他顶复门寄生虫的生长。在本申请中，我们将利用弓形虫的遗传和实验易处理性，从疟疾盒收集中鉴定出阻断弓形虫和疟原虫生长的化合物。然后，我们将使用化学诱变来分离耐药弓形虫寄生虫，并确定赋予弓形虫耐药性的突变。然后将在恶性疟原虫中检查耐药等位基因，以确定哪些耐药基因是保守的，从而代表泛顶复门药物靶标。

DESCRIPTION (provided by applicant): Apicomplexan parasites include important human pathogens such as Plasmodium spp., Toxoplasma gondii, and Cryptosporidium parvum. Due to a lack of vaccines as well as continuously developing resistance and severe side-effects to currently available treatments, new drugs are needed to treat these infections. While high-throughput small molecule screening has identified numerous anti-malarial lead compounds, it is not known how almost all of these compounds kill Plasmodium. Understanding these mechanisms is important for two key reasons. First, developing a compound into a drug that can be prescribed to patients requires a thorough understanding of the compound's mechanism of action. Second, drugs work by inhibiting cellular compounds and pathways and discovering what these are reveals new drug targets. Because many Apicomplexan-specific processes are essential for growth of these parasites, it is likely that some of lead anti-malarial lead compounds will also affect growth of other Apicomplexan parasites. In this application, we will take advantage of the genetic and experimental tractability of Toxoplasma gondii to identify compounds from the Malaria Box collection that block growth of both Toxoplasma and Plasmodium. We will then use chemical mutagenesis to isolate drug resistant Toxoplasma parasites and identify the mutations that confer drug resistance in Toxoplasma. The drug resistance alleles will then be examined in Plasmodium falciparum to determine which drug resistance genes are conserved and thus represent pan-Apicomplexan drug targets.