Summary of Study ST003144
This data is 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 Project ID PR001955. The data can be accessed directly via it's Project DOI: 10.21228/M8172C This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST003144 |
| Study Title | On-target, dual aminopeptidase inhibition provides cross-species antimalarial activity |
| Study Summary | To combat the global burden of malaria, development of new drugs to replace or complement current therapies are 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 haemoglobin digestion in asexual parasites. MMV1557817 can kill sexual stage P. falciparum, is active against murine malaria and did 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 sensitised to the current clinical drug, artemisinin. Overall, these results confirm MMV1557817 as a 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. |
| Institute | Monash University |
| Last Name | Siddiqui |
| First Name | Ghizal |
| Address | 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia |
| ghizal.siddiqui@monash.edu | |
| Phone | 99039282 |
| Submit Date | 2024-03-25 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-04-18 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001955 |
| Project DOI: | doi: 10.21228/M8172C |
| Project Title: | On-target, dual aminopeptidase inhibition provides cross-1 species antimalarial activity |
| Project Summary: | To combat the global burden of malaria, development of new drugs to replace or complement current therapies are 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 haemoglobin digestion in asexual parasites. MMV1557817 can kill sexual stage P. falciparum, is active against murine malaria and did 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 sensitised to the current clinical drug, artemisinin. Overall, these results confirm MMV1557817 as a 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. |
| Institute: | Monash University |
| Last Name: | Siddiqui |
| First Name: | Ghizal |
| Address: | 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia |
| Email: | ghizal.siddiqui@monash.edu |
| Phone: | 99039282 |
Subject:
| Subject ID: | SU003261 |
| Subject Type: | Cultured cells |
| Subject Species: | Plasmodium falciparum |
| Taxonomy ID: | 5833 |
Factors:
Subject type: Cultured cells; Subject species: Plasmodium falciparum (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | treatment |
|---|---|---|---|
| SA340687 | 78_irbcs_p_4 | iRBC | Compound MMV1557817 |
| SA340688 | 78_irbcs_p_3 | iRBC | Compound MMV1557817 |
| SA340689 | 78_irbcs_p_2 | iRBC | Compound MMV1557817 |
| SA340690 | 78_irbcs_p_1 | iRBC | Compound MMV1557817 |
| Showing results 1 to 4 of 4 |
Collection:
| Collection ID: | CO003254 |
| Collection Summary: | Metabolomics analysis of MMV1557817, MIPS2673 (ST002792) and 3 compared to DMSO control (ST002108): P. falciparum (3D7) cultures at 6% parasitaemia and 2% haematocrit were subject to double sorbitol synchronisation 14 hours apart, followed by further incubation for 28-42 h to achieve the desired trophozoite stage (28 hours post infection). Infected RBCs (2x108) were treated with 10x the EC50 of MMV1557817 (320 nM), MIPS2673 (1 µM), 3 (4.53 µM) for 1 h, after which metabolites were extracted, analysed and processed as previously described (7). Principal-component analysis (PCA) and hierarchical clustering algorithms were run also in Metaboanalyst (50). Metabolomics data are presented as relative abundance values from 4-7 biological replicates. Differences were determined using Welch’s t-test where significant interactions were observed. Significance was determined at p values < 0.05. The metabolomics data for 3, MIPS-2673 and drug-free controls were reported previously (7, 51). |
| Sample Type: | Blood (whole) |
Treatment:
| Treatment ID: | TR003270 |
| Treatment Summary: | Metabolomics analysis of MMV1557817, MIPS2673 (ST002792) and 3 compared to DMSO control (ST002108): P. falciparum (3D7) cultures at 6% parasitaemia and 2% haematocrit were subject to double sorbitol synchronisation 14 hours apart, followed by further incubation for 28-42 h to achieve the desired trophozoite stage (28 hours post infection). Infected RBCs (2x108) were treated with 10x the EC50 of MMV1557817 (320 nM), MIPS2673 (1 µM), 3 (4.53 µM) for 1 h, after which metabolites were extracted, analysed and processed as previously described (7). Principal-component analysis (PCA) and hierarchical clustering algorithms were run also in Metaboanalyst (50). Metabolomics data are presented as relative abundance values from 4-7 biological replicates. Differences were determined using Welch’s t-test where significant interactions were observed. Significance was determined at p values < 0.05. The metabolomics data for 3, MIPS-2673 and drug-free controls were reported previously (7, 51). |
Sample Preparation:
| Sampleprep ID: | SP003268 |
| Sampleprep Summary: | Metabolomics analysis of MMV1557817, MIPS2673 (ST002792) and 3 compared to DMSO control (ST002108): P. falciparum (3D7) cultures at 6% parasitaemia and 2% haematocrit were subject to double sorbitol synchronisation 14 hours apart, followed by further incubation for 28-42 h to achieve the desired trophozoite stage (28 hours post infection). Infected RBCs (2x108) were treated with 10x the EC50 of MMV1557817 (320 nM), MIPS2673 (1 µM), 3 (4.53 µM) for 1 h, after which metabolites were extracted, analysed and processed as previously described (7). Principal-component analysis (PCA) and hierarchical clustering algorithms were run also in Metaboanalyst (50). Metabolomics data are presented as relative abundance values from 4-7 biological replicates. Differences were determined using Welch’s t-test where significant interactions were observed. Significance was determined at p values < 0.05. The metabolomics data for 3, MIPS-2673 and drug-free controls were reported previously (7, 51). |
Combined analysis:
| Analysis ID | AN005159 | AN005160 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo | Thermo |
| Column | Merck SeQuant ZIC-pHILIC (150 x 4.6mm,5um) | Merck SeQuant ZIC-pHILIC (150 x 4.6mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peak height | peak height |
Chromatography:
| Chromatography ID: | CH003905 |
| Instrument Name: | Thermo |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 4.6mm,5um) |
| Column Temperature: | 25 |
| Flow Gradient: | 0% B to 50% B over 15 min, then to 5% B at 18 min until 21 min, increasing to 80% B at 24 min until 32 min. |
| Flow Rate: | 0.3 ml/min |
| Solvent A: | 100% water; 20 mM ammonium carbonate |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004895 |
| Analysis ID: | AN005159 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Metabolite detection was performed using a high-resolution Q Exactive MS (ThermoFisher) in both positive and negative ionisation modes. The PBQC sample was run periodically throughout each LC-MS batch to monitor signal reproducibility and support downstream metabolite identification. Extraction solvent blank samples were also analysed to identify possible contaminating chemical species. To aid in metabolite identification, approximately 250 authentic metabolite standards were analysed prior to each LC-MS batch and their peaks and retention time manually checked using the ToxID software (ThermoFisher). Metabolomics data were analysed using the IDEOM workflow (Creek et al. 2012). Briefly, the IDEOM processing pipeline uses msconvert for conversion of raw files to mzXML files and split polarity, XCMS to extract raw peak intensities and mzMatch to align samples, filter noise, fill missing peaks and annotate related peaks. Manual assessment of spiked internal standards, total ion chromatograms and median peak heights ensured signal reproducibility and allowed exclusion of outlier samples. LC MS peak heights representing metabolite abundances were normalised by median peak height. High confidence metabolite identification (MSI level 1) was made by matching accurate mass and retention time to authentic metabolite standards. Putative identifications (MSI level 2) for metabolites lacking standards were based on exact mass and predicted retention times. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004896 |
| Analysis ID: | AN005160 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Metabolite detection was performed using a high-resolution Q Exactive MS (ThermoFisher) in both positive and negative ionisation modes. The PBQC sample was run periodically throughout each LC-MS batch to monitor signal reproducibility and support downstream metabolite identification. Extraction solvent blank samples were also analysed to identify possible contaminating chemical species. To aid in metabolite identification, approximately 250 authentic metabolite standards were analysed prior to each LC-MS batch and their peaks and retention time manually checked using the ToxID software (ThermoFisher). Metabolomics data were analysed using the IDEOM workflow (Creek et al. 2012). Briefly, the IDEOM processing pipeline uses msconvert for conversion of raw files to mzXML files and split polarity, XCMS to extract raw peak intensities and mzMatch to align samples, filter noise, fill missing peaks and annotate related peaks. Manual assessment of spiked internal standards, total ion chromatograms and median peak heights ensured signal reproducibility and allowed exclusion of outlier samples. LC MS peak heights representing metabolite abundances were normalised by median peak height. High confidence metabolite identification (MSI level 1) was made by matching accurate mass and retention time to authentic metabolite standards. Putative identifications (MSI level 2) for metabolites lacking standards were based on exact mass and predicted retention times. |
| Ion Mode: | NEGATIVE |
