Summary of Study ST003160
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 PR001965. The data can be accessed directly via it's Project DOI: 10.21228/M8QQ8P 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 | ST003160 |
| Study Title | New class of heterospirocyclic compounds present strong and rapid activity against artemisinin- and multidrug-resistant P. falciparum parasites |
| Study Summary | Malaria remains a significant health burden and a leading contributor to global mortality rates. Increasing drug resistance creates an urgent demand for novel treatment options. We have synthesised a new class of heterospirocyclic compounds with novel chemical connectivities. Compounds 25 and 26 display antimalarial activity within 24 h and have similar potency against a panel of drug-resistant strains of Plasmodium falciparum, the most virulent of human malaria parasites, including parasites resistant to the frontline artemisinin antimalarials. C25 and C26 do not induce major toxicity in kidney- and hepatic-derived human cell lines, highlighting their specificity. Untargeted metabolomics analysis of P. falciparum infected red blood cells revealed that the mechanism of action of C25 involves disruption of the pyrimidine biosynthesis pathway and haemoglobin catabolism. These heterospirocyclic compounds represent a promising opportunity for antimalarial drug development and could prove relevant against drug resistant malaria. |
| Institute | Monash University |
| Last Name | Giannangelo |
| First Name | Carlo |
| Address | 381 Royal Parade, Parkville, Victoria, 3052, Australia |
| carlo.giannangelo@monash.edu | |
| Phone | 99039282 |
| Submit Date | 2024-04-06 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-04-29 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001965 |
| Project DOI: | doi: 10.21228/M8QQ8P |
| Project Title: | New class of heterospirocyclic compounds present strong and rapid activity against artemisinin- and multidrug-resistant P. falciparum parasites |
| Project Summary: | Malaria remains a significant health burden and a leading contributor to global mortality rates. Increasing drug resistance creates an urgent demand for novel treatment options. We have synthesised a new class of heterospirocyclic compounds with novel chemical connectivities. Compounds 25 and 26 display antimalarial activity within 24 h and have similar potency against a panel of drug-resistant strains of Plasmodium falciparum, the most virulent of human malaria parasites, including parasites resistant to the frontline artemisinin antimalarials. C25 and C26 do not induce major toxicity in kidney- and hepatic-derived human cell lines, highlighting their specificity. Untargeted metabolomics analysis of P. falciparum infected red blood cells revealed that the mechanism of action of C25 involves disruption of the pyrimidine biosynthesis pathway and haemoglobin catabolism. These heterospirocyclic compounds represent a promising opportunity for antimalarial drug development and could prove relevant against drug resistant malaria. |
| Institute: | Monash University |
| Last Name: | Giannangelo |
| First Name: | Carlo |
| Address: | 381 Royal Parade, Parkville, Victoria, 3052, Australia |
| Email: | carlo.giannangelo@monash.edu |
| Phone: | 99039282 |
Subject:
| Subject ID: | SU003279 |
| Subject Type: | Cultured cells |
| Subject Species: | Plasmodium falciparum |
| Taxonomy ID: | 5833 |
| Species Group: | Unicellular parasites |
Factors:
Subject type: Cultured cells; Subject species: Plasmodium falciparum (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA342802 | ATV_1 | Atovaquone |
| SA342803 | ATV_4 | Atovaquone |
| SA342804 | ATV_3 | Atovaquone |
| SA342805 | ATV_2 | Atovaquone |
| SA342806 | C25_4 | C25 |
| SA342807 | C25_3 | C25 |
| SA342808 | C25_1 | C25 |
| SA342809 | C25_2 | C25 |
| SA342810 | C25_0HR_3 | C25_0HR_control |
| SA342811 | C25_0HR_2 | C25_0HR_control |
| SA342812 | C25_0HR_4 | C25_0HR_control |
| SA342813 | C25_0HR_1 | C25_0HR_control |
| SA342814 | DMSO_3 | DMSO |
| SA342815 | DMSO_2 | DMSO |
| SA342816 | DMSO_1 | DMSO |
| SA342817 | DMSO_4 | DMSO |
| SA342818 | PS027_1 | PS027 |
| SA342819 | PS027_4 | PS027 |
| SA342820 | PS027_2 | PS027 |
| SA342821 | PS027_3 | PS027 |
| Showing results 1 to 20 of 20 |
Collection:
| Collection ID: | CO003272 |
| Collection Summary: | Plasmodium falciparum cultures (3D7 strain) synchronised to the trophozoite stage were magnet purified to achieve a parasitaemia of >90% and haematocrit of 0.5%. Four different experiment conditions were used: (i) the active heterospirocyclic compound, compound 25 (C25); (ii) the inactive precursor of compound 25, PS027; (iii) atovaquone; and (iv) DMSO alone as the negative (no drug) control. Following compound incubation (5 h), cultures were centrifuged at 2,000 RPM for 5 minutes, the media was removed and the cell pellets were washed in ice cold PBS. All samples were centrifuged at 1,200g for 3 minutes, after which metabolites were extracted from 5e7 cells using 100 µL of ice-cold methanol extraction solvent. Samples were placed on a shaker at 4°C for 1 h and spun at 13,000 RPM for 10 minutes at 4 °C. Supernatants were transferred into high-performance liquid chromatography (HPLC) vials and stored at -80°C until liquid chromatography-mass spectrometry (LC-MS) analysis. A pooled biological quality control sample consisting of a 15 µL aliquot from each sample was also generated for analytical quality control and to aid metabolite identification. |
| Sample Type: | Plasmodium cells |
Treatment:
| Treatment ID: | TR003288 |
| Treatment Summary: | Plasmodium falciparum cultures (3D7 strain) synchronised to the trophozoite stage were magnet purified to achieve a parasitaemia of >90% and haematocrit of 0.5%. Four different experiment conditions were used: (i) the active heterospirocyclic compound, compound 25 (C25); (ii) the inactive precursor of compound 25, PS027; (iii) atovaquone; and (iv) DMSO alone as the negative (no drug) control. Following compound incubation (5 h), cultures were centrifuged at 2,000 RPM for 5 minutes, the media was removed and the cell pellets were washed in ice cold PBS. All samples were centrifuged at 1,200g for 3 minutes, after which metabolites were extracted from 5e7 cells using 100 µL of ice-cold methanol extraction solvent. Samples were placed on a shaker at 4°C for 1 h and spun at 13,000 RPM for 10 minutes at 4 °C. Supernatants were transferred into high-performance liquid chromatography (HPLC) vials and stored at -80°C until liquid chromatography-mass spectrometry (LC-MS) analysis. A pooled biological quality control sample consisting of a 15 µL aliquot from each sample was also generated for analytical quality control and to aid metabolite identification. |
Sample Preparation:
| Sampleprep ID: | SP003286 |
| Sampleprep Summary: | Plasmodium falciparum cultures (3D7 strain) synchronised to the trophozoite stage were magnet purified to achieve a parasitaemia of >90% and haematocrit of 0.5%. Four different experiment conditions were used: (i) the active heterospirocyclic compound, compound 25 (C25); (ii) the inactive precursor of compound 25, PS027; (iii) atovaquone; and (iv) DMSO alone as the negative (no drug) control. Following compound incubation (5 h), cultures were centrifuged at 2,000 RPM for 5 minutes, the media was removed and the cell pellets were washed in ice cold PBS. All samples were centrifuged at 1,200g for 3 minutes, after which metabolites were extracted from 5e7 cells using 100 µL of ice-cold methanol extraction solvent. Samples were placed on a shaker at 4°C for 1 h and spun at 13,000 RPM for 10 minutes at 4 °C. Supernatants were transferred into high-performance liquid chromatography (HPLC) vials and stored at -80°C until liquid chromatography-mass spectrometry (LC-MS) analysis. A pooled biological quality control sample consisting of a 15 µL aliquot from each sample was also generated for analytical quality control and to aid metabolite identification. |
Chromatography:
| Chromatography ID: | CH003922 |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Merck SeQuant ZIC-HILIC (150 x 4.6mm,5um) |
| Column Temperature: | 25 |
| Flow Gradient: | 0–15 min, 80–50% B; 15–18 min, 50–5% B; 18–21 min, 5% B; 21–24 min, 5–80% B and 24–32 min, 80% B |
| Flow Rate: | 0.3 mL/min |
| Solvent A: | 100% Water; 20 mM ammonium carbonate |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN005184 |
| Analysis Type: | MS |
| Chromatography ID: | CH003922 |
| Num Factors: | 5 |
| Num Metabolites: | 341 |
| Rt Units: | Minutes |
| Units: | Peak height |
| Analysis ID: | AN005185 |
| Analysis Type: | MS |
| Chromatography ID: | CH003922 |
| Num Factors: | 5 |
| Num Metabolites: | 401 |
| Rt Units: | Minutes |
| Units: | Peak height |
