The malaria parasite Plasmodium obligatorily infects and replicates inside hepatocytes surrounded by a parasitophorous vacuole membrane (PVM), which is decorated by the host-cell derived autophagy protein LC3. We have previously shown that the parasite-derived, PVM-resident protein UIS3 sequesters LC3 to avoid parasite elimination by autophagy from hepatocytes. Here we show that a small molecule capable of disrupting this interaction triggers parasite elimination in a host cell autophagy-dependent manner. Molecular docking analysis of more than 20 million compounds combined with a phenotypic screen identified one molecule, C4 (4-{[4-(4-{5-[3-(trifluoromethyl) phenyl]-1,2,4-oxadiazol-3-yl}benzyl)piperazino]carbonyl}benzonitrile), capable of impairing infection. Using biophysical assays, we established that this impairment is due to the ability of C4 to disrupt UIS3–LC3 interaction, thus inhibiting the parasite’s ability to evade the host autophagy response. C4 impacts infection in autophagy-sufficient cells without harming the normal autophagy pathway of the host cell. This study, by revealing the disruption of a critical host–parasite interaction without affecting the host’s normal function, uncovers an efficient anti-malarial strategy to prevent this deadly disease.

疟原虫疟原虫专门感染并复制被寄生虫空泡膜（PVM）包围的肝细胞内部，该膜由宿主细胞衍生的自噬蛋白LC3修饰。先前我们已经表明，寄生虫衍生的PVM驻留蛋白UIS3隔离LC3，以避免通过自噬从肝细胞中消除寄生虫。在这里，我们表明能够破坏这种相互作用的小分子以宿主细胞自噬依赖性方式触发寄生虫消除。结合表型筛选对超过2000万种化合物进行分子对接分析，鉴定出一个分子C4（4-{[4-（4- {5- [3- [3-（三氟甲基）苯基] -1,2,4-恶二唑-3 -基}苄基）哌嗪子基]羰基}苄腈），能够削弱感染。使用生物物理分析，我们确定这种损伤是由于C4破坏UIS3-LC3相互作用的能力，从而抑制了寄生虫逃避宿主自噬反应的能力。C4影响自噬充足细胞中的感染，而不会损害宿主细胞的正常自噬途径。这项研究通过揭示关键的宿主-寄生虫相互作用的破坏而不影响宿主的正常功能，揭示了一种预防这种致命疾病的有效抗疟疾策略。