Abstract:
New antimalarial drug candidates that act via novel mechanisms are urgently needed to combat malaria drug resistance. Here, we describe the multi-omic chemical validation of Plasmodium M1 alanyl metalloaminopeptidase as an attractive drug target using the selective inhibitor, MIPS2673. MIPS2673 demonstrated potent inhibition of recombinant Plasmodium falciparum (PfA-M1) and Plasmodium vivax (PvA-M1) M1 metalloaminopeptidases, with selectivity over other Plasmodium and human aminopeptidases, and displayed excellent in vitro antimalarial activity with no significant host cytotoxicity. Orthogonal label-free chemoproteomic methods based on thermal stability and limited proteolysis of whole parasite lysates revealed that MIPS2673 solely targets PfA-M1 in parasites, with limited proteolysis also enabling estimation of the binding site on PfA-M1 to within ~5 Å of that determined by X-ray crystallography. Finally, functional investigation by untargeted metabolomics demonstrated that MIPS2673 inhibits the key role of PfA-M1 in haemoglobin digestion. Combined, our unbiased multi-omic target deconvolution methods confirmed the on-target activity of MIPS2673, and validated selective inhibition of M1 alanyl metalloaminopeptidase as a promising antimalarial strategy.
摘要:
迫切需要通过新机制起作用的新的抗疟疾候选药物来对抗疟疾耐药性。这里,我们描述了使用选择性抑制剂作为有吸引力的药物靶标的疟原虫M1丙氨酰金属氨基肽酶的多组化学验证,MIPS2673。MIPS2673显示了对重组恶性疟原虫(PfA-M1)和间日疟原虫(PvA-M1)M1金属氨基肽酶的有效抑制,具有对其他疟原虫和人类氨肽酶的选择性,并显示出优异的体外抗疟活性,没有明显的宿主细胞毒性。基于热稳定性和整个寄生虫裂解物的有限蛋白水解的正交无标记化学蛋白质组学方法表明,MIPS2673仅靶向寄生虫中的PfA-M1,有限的蛋白水解还可以估计PfA-M1上的结合位点,使其在X射线晶体学测定的范围内。最后,非靶向代谢组学的功能研究表明,MIPS2673抑制PfA-M1在血红蛋白消化中的关键作用.合并,我们的无偏多组目标去卷积方法证实了MIPS2673的中靶活性,并验证了M1丙氨酰金属氨基肽酶的选择性抑制是一种有前景的抗疟策略.