Chemoproteomics validates selective targeting of Plasmodium M1 alanyl aminopeptidase as a cross-species strategy to treat malaria

Creek, Darren; Giannangelo, Carlo; Challis, Matthew; Siddiqui, Ghizal; Edgar, Rebecca; Malcolm, Tess; Webb, Chaille; Drinkwater, Nyssa; Vinh, Natalie; MacRaild, Christopher; Counihan, Natalie; Duffy, Sandra; Wittlin, Sergio; Devine, Shane; Avery, Vicky; de Koning-Ward, Tania; Scammells, Peter; McGowan, Sheena

doi:10.21203/rs.3.rs-3251230/v1

Cited by 2 publications

(9 citation statements)

References 86 publications

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“…Value represents the mean of two biological replicates performed with ≥3 technical replicates ± standard deviation. ( C ) Hierarchical clustering of the 73 significantly perturbed peptides ( P value < 0.05) following treatment with MMV1557817 , MIPS2673 ( Pf A-M1 inhibitor [ 53 ]), 3 ( Pf A-M17 inhibitor [ 7 ]), and DMSO control. Vertical clustering displays similarities between samples, while horizontal clusters reveal the relative abundances of the 73 peptides from four to seven biological replicates.…”

Section: Resultsmentioning

confidence: 99%

“…To further confirm that MMV1557817 targets Pf A-M1 and Pf A-M17, we compared metabolomics profiles of Pf 3D7 parasites treated with 10× EC 50 of MMV1557817 , MIPS2673 ( Pf A-M1 inhibitor [ 53 ]), 3 ( Pf A-M17 inhibitor [ 7 ]), or DMSO control (Fig. S3).…”

Section: Resultsmentioning

confidence: 99%

“…4C ), consistent with additive inhibition of both aminopeptidases. We have previously shown that majority of these peptides could be mapped to hemoglobin sequences ( 7 , 53 ).…”

Section: Resultsmentioning

confidence: 99%

“…Significance was determined at P values < 0.05. The metabolomics data for 3 , MIPS2673, and drug-free controls were reported previously ( 7 , 53 ). The metabolomics data are available at the NIH Common Fund’s National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org where it has been assigned Study ID ST003144.…”

Section: Methodsmentioning

confidence: 99%

“…After 10 min incubation at 37°C, 10 µL of treated parasites was spotted onto a glass slide, covered with a coverslip, and imaged using a 4′,6-diamidino-2-phenylindole (DAPI) filter set on a Nikon Eclipse Ti2 microscope at 100× magnification under oil immersion. To inhibit Pf A-M1 activity, washed parasites were treated with MIPS2673 (3.2 µM; 10× EC 50 ) ( 53 ) for 20 min prior to substrate addition. The relative fluorescence detected within a parasite cytosol was calculated using ImageJ (NIH, version 1.53c) and was expressed relative to the surface area.…”

Section: Methodsmentioning

confidence: 99%

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On-target, dual aminopeptidase inhibition provides cross-species antimalarial activity

Edgar,

Malcolm,

Siddiqui

et al. 2024

mBio

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To combat the global burden of malaria, development of new drugs to replace or complement current therapies is urgently required. Here, we show that the compound MMV1557817 is a selective, nanomolar inhibitor of both Plasmodium falciparum and Plasmodium vivax aminopeptidases M1 and M17, leading to inhibition of end-stage hemoglobin digestion in asexual parasites. MMV1557817 can kill sexual-stage P. falciparum , is active against murine malaria, and does not show any shift in activity against a panel of parasites resistant to other antimalarials. MMV1557817 -resistant P. falciparum exhibited a slow growth rate that was quickly outcompeted by wild-type parasites and were sensitized to the current clinical drug, artemisinin. Overall, these results confirm MMV1557817 as a lead compound for further drug development and highlights the potential of dual inhibition of M1 and M17 as an effective multi-species drug-targeting strategy. IMPORTANCE Each year, malaria infects approximately 240 million people and causes over 600,000 deaths, mostly in children under 5 years of age. For the past decade, artemisinin-based combination therapies have been recommended by the World Health Organization as the standard malaria treatment worldwide. Their widespread use has led to the development of artemisinin resistance in the form of delayed parasite clearance, alongside the rise of partner drug resistance. There is an urgent need to develop and deploy new antimalarial agents with novel targets and mechanisms of action. Here, we report a new and potent antimalarial compound, known as MMV1557817 , and show that it targets multiple stages of the malaria parasite lifecycle, is active in a preliminary mouse malaria model, and has a novel mechanism of action. Excitingly, resistance to MMV15578117 appears to be self-limiting, suggesting that development of the compound may provide a new class of antimalarial.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…4C ), consistent with additive inhibition of both aminopeptidases. We have previously shown that majority of these peptides could be mapped to hemoglobin sequences ( 7 , 53 ).…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

On-target, dual aminopeptidase inhibition provides cross-species antimalarial activity

Edgar,

Malcolm,

Siddiqui

et al. 2024

mBio

Self Cite

View full text Add to dashboard Cite

show abstract

On-target, dual aminopeptidase inhibition provides cross-species antimalarial activity

Edgar,

Malcolm,

Siddiqui

et al. 2023

Preprint

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To combat the global burden of malaria, development of new drugs to replace or complement current therapies are urgently required. As drug resistance to existing treatments and clinical failures continue to rise, compounds targeting multiple life cycle stages and species need to be developed as a high priority. Here we show that the compoundMMV1557817is a nanomolar inhibitor of bothPlasmodium falciparumandPlasmodium vivaxaminopeptidases M1 and M17, leading to inhibition of end stage haemoglobin digestion in asexual parasites. Multi-stage analysis confirmed thatMMV1557817can also kill sexual stageP. falciparum, while cross-resistance studies confirmed the compound targets a mechanism of action distinct to current drug resistance mechanisms. Analysis of cross reactivity to homologous human enzymes shows the compound exhibits a high level of selectivity, whilst safety as well as druggability was confirmed in the murine modelP. berghei.MMV1557817-resistantP. falciparumparasites displayed only low-level resistance (<3-fold) and exhibited a slow growth rate that was quickly outcompeted by wild type parasites.MMV1557817-resistant parasites digest significantly more haemoglobin and possess a mutation inPfA-M17 that induces partial destabilisation of thePfA-M17 homohexamer, resulting in high-level resistance to specificPfA-M17 inhibition, but enhanced sensitivity to specificPfA-M1 inhibition, and importantly, these parasites were highly sensitive to artemisinin. Overall, these results confirmMMV1557817as a potential lead compound for further drug development and highlight the potential of dual inhibition of M1 and M17 as an effective multi-species drug targeting strategy.

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Chemoproteomics validates selective targeting of Plasmodium M1 alanyl aminopeptidase as a cross-species strategy to treat malaria

Cited by 2 publications

References 86 publications

On-target, dual aminopeptidase inhibition provides cross-species antimalarial activity

On-target, dual aminopeptidase inhibition provides cross-species antimalarial activity

On-target, dual aminopeptidase inhibition provides cross-species antimalarial activity

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