Center for Malaria Therapeutics and Antimicrobial Resistance and Department of Microbiology and Immunology, Columbia University Irving Medical Center, New York, NY, USA.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Cell Chem Biol. 2023 May 18;30(5):470-485.e6. doi: 10.1016/j.chembiol.2023.03.002. Epub 2023 Mar 23.
The Plasmodium falciparum proteasome constitutes a promising antimalarial target, with multiple chemotypes potently and selectively inhibiting parasite proliferation and synergizing with the first-line artemisinin drugs, including against artemisinin-resistant parasites. We compared resistance profiles of vinyl sulfone, epoxyketone, macrocyclic peptide, and asparagine ethylenediamine inhibitors and report that the vinyl sulfones were potent even against mutant parasites resistant to other proteasome inhibitors and did not readily select for resistance, particularly WLL that displays covalent and irreversible binding to the catalytic β2 and β5 proteasome subunits. We also observed instances of collateral hypersensitivity, whereby resistance to one inhibitor could sensitize parasites to distinct chemotypes. Proteasome selectivity was confirmed using CRISPR/Cas9-edited mutant and conditional knockdown parasites. Molecular modeling of proteasome mutations suggested spatial contraction of the β5 P1 binding pocket, compromising compound binding. Dual targeting of P. falciparum proteasome subunits using covalent inhibitors provides a potential strategy for restoring artemisinin activity and combating the spread of drug-resistant malaria.
疟原虫蛋白酶体是一种很有前途的抗疟靶点，多种化学型能够强效且选择性地抑制寄生虫的增殖，并与一线青蒿素类药物协同作用，包括对耐青蒿素的寄生虫。我们比较了乙烯砜、环氧化酮、大环肽和天冬酰胺乙二胺抑制剂的耐药谱，并报告说乙烯砜即使对其他蛋白酶体抑制剂耐药的突变寄生虫也具有很强的抑制作用，而且不易产生耐药性，特别是 WLL 与催化β2 和β5 蛋白酶体亚基发生共价和不可逆结合。我们还观察到一些偶联的超敏现象，即对一种抑制剂的耐药性会使寄生虫对不同的化学型敏感。使用 CRISPR/Cas9 编辑突变体和条件性敲低寄生虫证实了蛋白酶体的选择性。蛋白酶体突变的分子建模表明β5 P1 结合口袋的空间收缩，损害了化合物的结合。使用共价抑制剂双重靶向疟原虫蛋白酶体亚基为恢复青蒿素活性和对抗耐药性疟疾的传播提供了一种潜在策略。
