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BioAssay: AID 488774

Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid: Summary

The goal of this screen is to discover new antimalarial compounds that act by inhibiting the development of the apicoplast in the malarial parasite Plasmodium falciparum. The biochemical processes that make this organelle essential for erythrocytic stage parasites are not well understood. However, antibiotics, such as azithromycin and tetracycline, which target the apicoplast translational more ..
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 Related BioAssays
AID: 488774
Data Source: NCGC (DDS001)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-10-12
Modify Date: 2011-04-19
Related Experiments
Show more
AIDNameTypeComment
488745Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid, 96 hour incubationConfirmatorydepositor-specified cross reference: LOPAC validation at 96 hr incubation
488752Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid, 48 hour incubationConfirmatorydepositor-specified cross reference: LOPAC validation at 48 hr incubation
504599Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid, 96 hour incubation.Confirmatorydepositor-specified cross reference: 96 hour incubation
504602Quantitative high throughput screen to test cell viability of anti-malarial compounds targeting the delayed death phenotypeConfirmatorydepositor-specified cross reference: Cell Viability
504604Quantitative high throughput screen to test activity of anti-malarial compounds in the Dd2 strain of Plasmodium FalciparumConfirmatorydepositor-specified cross reference: P. Falciparum activity
504608Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid, 48 hour incubation.Confirmatorydepositor-specified cross reference: 48 hour incubation
504631A cell-based HTS for delayed death inhibitors of the malarial parasite plastid Measured in Microorganism System Using Plate Reader - 2126-02_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference: Broad Institute Assay
504633A cell-based HTS for delayed death inhibitors of the malarial parasite plastid Measured in Microorganism System Using Plate Reader - 2126-01_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference: Broad Institute Assay
504658FACS-based assay to assess parasite growth inhibition 48 hr Measured in Cell-Based System Using Flow Cytometry - 2126-03_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference: Broad Institute Assay
504659FACS-based assay to assess parasite growth inhibition 96 hr Measured in Cell-Based System Using Flow Cytometry - 2126-05_Inhibitor_Dose_DryPowder_ActivityConfirmatorydepositor-specified cross reference: Broad Institute Assay
504661Cell-based assay to measure Apicoplast Disruption using ImagingOtherdepositor-specified cross reference: Broad Institute Assay
504832Primary qHTS for delayed death inhibitors of the malarial parasite plastid, 48 hour incubationConfirmatorydepositor-specified cross reference
504834Primary qHTS for delayed death inhibitors of the malarial parasite plastid, 96 hour incubationConfirmatorydepositor-specified cross reference
624206Quantitative high throughput screen to test cell viability of anti-malarial compounds targeting the delayed death phenotype.Confirmatorydepositor-specified cross reference
624328Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid: Hit Validation at 48 hrConfirmatorydepositor-specified cross reference
624329Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid: Hit Validation with 3D7 at 48 hr using Flow CytometryConfirmatorydepositor-specified cross reference
624332Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid: Hit Validation at 96 hrConfirmatorydepositor-specified cross reference
624335Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid: Hit Validation with 3D7 at 96 hr using Flow CytometryConfirmatorydepositor-specified cross reference
624336Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid: Hit Validation with Dd2 at 48 hr using Flow CytometryConfirmatorydepositor-specified cross reference
624337Quantitative high throughput screen for delayed death inhibitors of the malarial parasite plastid: Hit Validation with Dd2 at 96hr using Flow CytometryConfirmatorydepositor-specified cross reference
Description:
NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]

MLPCN Grant: R21 NS059500
Assay Provider: David Fidock and Eric Ekland, Columbia University

The goal of this screen is to discover new antimalarial compounds that act by inhibiting the development of the apicoplast in the malarial parasite Plasmodium falciparum. The biochemical processes that make this organelle essential for erythrocytic stage parasites are not well understood. However, antibiotics, such as azithromycin and tetracycline, which target the apicoplast translational machinery, have a potent antimalarial effect. The killing caused by these drugs is unusual in that it does not appear to affect the first generation of parasites that are exposed to the drug, but rather manifests itself in their progeny.

We have developed a cell-based assay that measures parasite growth based on the expression of an integrated copy of a firefly luciferase reporter. To detect small molecules that cause this 'delayed death' phenotype, erythrocytes infected with the luciferase-expressing parasites were incubated with compounds for either one or two generations, corresponding to 48 and 96 hours, respectively. Compounds that inhibit parasite growth in the second generation, but not the first, should be enriched in antimalarials that target the apicoplast. Growth inhibition is detected by a decrease in luciferase activity
Keywords: NIH Roadmap, MLPCN, MLI, MLSMR, qHTS, NCGC, qHTS, malaria, luciferase, viability
Protocol
For the 48 hour time point, the protocol is as follows:

Four microliters of culture medium (RPMI 1640 with 0.5% w/v Albumax (GIBCO), 24 mM sodium bicarbonate and 10 ug/mL gentamycin) were dispensed by a Multi-drop Combi into white solid 1536-well plates (Grenier) and 23 nL compound was added by a pin tool. Four microliters of infected erythrocytes (2% hematocrit, 0.1% parasitemia final concentration) in culture medium were dispensed and the plates incubated for 48 hours at 37 C in 5% CO2. Two microliters of luciferase detection reagent was added and luminescence was detected by a ViewLux (PerkinElmer) reader

For the 96 hour time point, the protocol is as follows:

Four microliters of culture medium (RPMI 1640 with 0.5% w/v Albumax (GIBCO), 24 mM sodium bicarbonate and 10 ug/mL gentamycin) were dispensed by a Multi-drop Combi into white solid 1536-well plates (Grenier) and 23 nL compound was added by a pin tool. Four microliters of infected erythrocytes (2% hematocrit, 0.1% parasitemia final concentration) in culture medium were dispensed and the plates incubated for 96 hours at 37 C in 5% CO2. Two microliters of luciferase detection reagent was added and luminescence was detected by a ViewLux (PerkinElmer) reader
Comment
This summary is written for the purposes of summarizing the probe activities from the project. MLSCN probes are given a score of 100. Molecules in the prior art are given a score of 80. Other, less active molecules in the same chemical series as the probe molecules are given a score of 50. Molecules pending validation are given a score of 10. Inactive analogues from these series are given a score of 0. This project is currently at the assay validation stage.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 1
MLP Probe ML# for probe 1: ML238
PubChem Substance ID (SID) for probe 1: 113585210
PubChem Compound ID (CID) for probe 1: 49849912
Probe type for probe 1: Inhibitor
IC50/EC50 (nM) for probe 1: 13
Target for probe 1: Plasmodium falciparum Dd2
Disease relevance for probe 1: Malaria
Anti-target for probe 1: Mammalian Cell Cytotoxicity
Fold selectivity for probe 1: 2620
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK114716/ (ID: 2994556)
Grant number for probe 1: NS059500-01
NCBI Book chapter title for probe 1: ML238: An Antimalarial Small Molecule of a Unique Structural Class
Additional Information
Grant Number: R21 NS059500

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