Summary of Study ST003179
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 PR001978. The data can be accessed directly via it's Project DOI: 10.21228/M8214C 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 | ST003179 |
| Study Title | Property and Activity Refinement of Dihydroquinazolinone-3-carboxamides as Orally Efficacious Antimalarials that Target PfATP4 |
| Study Summary | The development of new antimalarial classes is pertinent because of resistance against the current antimalarial treatments. To contribute to the global effort to create new antimalarial therapies, we previously disclosed initial findings on the optimization of the dihydroquinazolinone-3-carboxamide class that targets PfATP4. The preliminary optimization generated analogs, such as 4, that exhibited potent in vitro asexual stage activity but only showed modest oral efficacy in a P. berghei mouse model attributed to its low aqueous solubility and modest metabolic stability. Here we report on correcting these parameters to improve in vivo efficacy. We show that the incorporation of heterocycle systems in the 8-position of the scaffold markedly improved aqueous solubility without a significant loss of asexual parasite activity. Certain configurations of pyrazoles in the 8-position were found to provide the greatest attainable balance between parasite activity, aqueous solubility, and metabolic stability. We were able to show that modifications made to the optimized analogs, such as WJM992 did not perturb the sensitivity to PfATP4 drug-resistant parasites or alter on-target activity in a PfATP4-associated parasite cytosolic Na+ flux assay and gave a distinct metabolic signature indicative of other PfATP4 inhibitors. The optimized analogs showed an appreciable efficacy in malaria mouse models and blocked sexual stage gamete development preventing transmission to mosquitoes. |
| 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 | 2024-04-23 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-05-15 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN005221 | AN005222 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| 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 Orbitrap Exploris 120 | Thermo Orbitrap Exploris 120 |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak height | Peak height |
MS:
| MS ID: | MS004954 |
| Analysis ID: | AN005221 |
| Instrument Name: | Thermo Orbitrap Exploris 120 |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data were acquired as a full scan in positive and negative ionization modes with a heated electrospray source and an Orbitrap resolution of 120,000 from 70 to 1,050 m/z. Ion source voltage was 3,500 V in positive mode and 2,500 V in negative mode. The ion transfer tube temperature was 325 °C and the vaporizer temperature was 350 °C. Gas mode was set to static with sheath gas, aux gas, and sweep gas at 50, 10, and 1, respectively. Samples within the LC-MS batch were sorted according to blocks of replicates and randomized. To facilitate metabolite identification, approximately 350 authentic metabolite standards were analyzed before the LC-MS batch, and their peaks and retention time were manually checked using the MZmine software. Pooled biological quality control samples and extraction solvent blanks were analyzed periodically throughout the batch to monitor LC-MS signal reproducibility and assist metabolite identification procedures. Raw LC-MS metabolomics data were analysed using the open source software, IDEOM (http://mzmatch.sourceforge.net/ideom.php). Briefly, the IDEOM workflow uses msconvert to convert raw files to mzXML format, XCMS (Centwave) to pick LC-MS peak signals, and MZmatch for alignment and annotation of related metabolite peaks. Default IDEOM parameters were used to eliminate unwanted noise and artifact peaks. Confident metabolite identification was made by matching accurate masses to the retention time of the ~350 authentic standards. When these authentic standards were unavailable, putative metabolite identification used accurate mass and predicted retention times, as previously described. Metabolite abundance was represented by LC-MS peak height. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004955 |
| Analysis ID: | AN005222 |
| Instrument Name: | Thermo Orbitrap Exploris 120 |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data were acquired as a full scan in positive and negative ionization modes with a heated electrospray source and an Orbitrap resolution of 120,000 from 70 to 1,050 m/z. Ion source voltage was 3,500 V in positive mode and 2,500 V in negative mode. The ion transfer tube temperature was 325 °C and the vaporizer temperature was 350 °C. Gas mode was set to static with sheath gas, aux gas, and sweep gas at 50, 10, and 1, respectively. Samples within the LC-MS batch were sorted according to blocks of replicates and randomized. To facilitate metabolite identification, approximately 350 authentic metabolite standards were analyzed before the LC-MS batch, and their peaks and retention time were manually checked using the MZmine software. Pooled biological quality control samples and extraction solvent blanks were analyzed periodically throughout the batch to monitor LC-MS signal reproducibility and assist metabolite identification procedures. Raw LC-MS metabolomics data were analysed using the open source software, IDEOM (http://mzmatch.sourceforge.net/ideom.php). Briefly, the IDEOM workflow uses msconvert to convert raw files to mzXML format, XCMS (Centwave) to pick LC-MS peak signals, and MZmatch for alignment and annotation of related metabolite peaks. Default IDEOM parameters were used to eliminate unwanted noise and artifact peaks. Confident metabolite identification was made by matching accurate masses to the retention time of the ~350 authentic standards. When these authentic standards were unavailable, putative metabolite identification used accurate mass and predicted retention times, as previously described. Metabolite abundance was represented by LC-MS peak height. |
| Ion Mode: | NEGATIVE |
