Chemoproteomics validates selective targeting of Plasmodium M1 alanyl aminopeptidase as an antimalarial strategy

• DOI: 10.21203/rs.3.rs-3251230/v2
• Publication Date: 2024-04-10T19:31:22Z
• Authors: Darren Creek; Carlo Giannangelo; Matthew Challis; Ghizal Siddiqui; Rebecca Edgar; Tess Malcolm; Chaille Webb; Nyssa Drinkwater; Natalie Vinh; Christopher MacRaild; Natalie Counihan; Sandra Duffy; Sergio Wittlin; Shane Devine; Vicky Avery; Tania de Koning-Ward; Peter Scammells; Sheena McGowan
• Keywords: Plasmodium (life cycle); Antimalarial Agent

<title>Abstract</title> New antimalarial drug candidates that act via novel mechanisms are urgently needed to combat malaria resistance. Here, we describe the multi-omic chemical validation of <italic>Plasmodium </italic>M1 alanyl metalloaminopeptidase as an attractive target using selective inhibitor, MIPS2673. MIPS2673 demonstrated potent inhibition recombinant falciparum</italic> (<italic>Pf</italic>A-M1) and vivax</italic> (<italic>Pv</italic>A-M1) M1 metalloaminopeptidases, with selectivity over other <italic>Plasmodium</italic> human aminopeptidases, displayed excellent <italic>in vitro</italic> activity no significant host cytotoxicity. Orthogonal label-free chemoproteomic methods based on thermal stability limited proteolysis whole parasite lysates revealed solely targets <italic>Pf</italic>A-M1 in parasites, also enabling estimation binding site within ~5 Å determined by X-ray crystallography. Finally, functional investigation untargeted metabolomics inhibits key role of<italic> Pf</italic>A-M1 haemoglobin digestion. Combined, our unbiased deconvolution confirmed on-target MIPS2673, validated a promising strategy.