qHTS for Inhibitors of binding or entry into cells for Marburg Virus
Marburg virus is a filovirus that is originated in Central and East Africa and can cause Marburg hemorrhagic fever in humans. Marburg virus is in the same taxonomic family as Ebola virus. Marburg virus is enveloped, nonsegmented, and contains a negative-sense RNA strand. To identify inhibitors of Marburg virus entry into cells, a pseudotype virus system was used where the nonpathogenic vesicular more ..
BioActive Compounds: 1038
Depositor Specified Assays
Marburg virus is a filovirus that is originated in Central and East Africa and can cause Marburg hemorrhagic fever in humans. Marburg virus is in the same taxonomic family as Ebola virus. Marburg virus is enveloped, nonsegmented, and contains a negative-sense RNA strand. To identify inhibitors of Marburg virus entry into cells, a pseudotype virus system was used where the nonpathogenic vesicular stomatitis virus (VSV) contains the Marburg virus envelop glycoproteins. This pseudotype virus, termed VSV-MARV, has a Photinus luciferase reporter gene inserted in its genome. For this assay, human embryonic kidney 293 cells are incubated with VSV-MARV and virus entry is detected by luminescence produced from the luciferase reporter. Small molecule inhibitors that block VSV-MARV binding or entry into cells are identified by a decrease of luciferase activity.
In a collaboration between University of Texas Medical Branch in Galveston, TX and the NIH Chemical Genomics Center (NCGC) a miniturized, high throughput, cell-based assay was developed and screened against the Molecular Libraries Small Molecule Repository (MLSMR) library.
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]
MLPCN Grant: MH086850-01A1
Assay Submitter (PI): Robert Davey, University of Texas Medical Branch, Galveston, TX
Two uL of HEK293 cell suspension are dispensed at 1000 cells/well into solid white 1536-well plates (Grenier) using a Multidrop Combi (Thermo Scientific). After addition of 23 nL compound by a pin tool (Kalypsys), the plate is incubated 1 h at 37 degrees C and then 3 uL of virus 1:100 dilution VSV-MARV is added. After 28 hr, 4 uL of assay reagent is added and the plates are read using a ViewLux (Perkin Elmer). Assays are performed in sub-saturating amounts of virus (MOI <0.5), therefore luciferase signals reflect the amount (titer) of virus able to infect the cells in presence of the compound.
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
* Activity Concentration. ** Test Concentration.
Data Table (Concise)