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BioAssay: AID 651812

qHTS for inhibitors of Vif-A3G interactions: Cherry picks

The recent discovery and characterization of host restriction factors, which provide a potent defense to invading retroviruses, has provided new targets for antiviral drug development. APOBEC3G (A3G) and APOBEC3F (A3F) are potent cytidine deaminases that block HIV-1 replication; HIV-1 expresses the Vif protein, which counteracts A3G and A3F by binding to them and inducing their proteasomal degradation. ..more
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 Tested Compounds
 Tested Compounds
All(2092)
 
 
Active(648)
 
 
Inactive(1109)
 
 
Inconclusive(336)
 
 
 Tested Substances
 Tested Substances
All(2094)
 
 
Active(649)
 
 
Inactive(1109)
 
 
Inconclusive(336)
 
 
AID: 651812
Data Source: NCGC (VIFG200)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-11-27

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 648
Depositor Specified Assays
AIDNameTypeComment
588444qHTS for inhibitors of Vif-A3G interactions: SummarysummaryVif-A3G Summary
602324qHTS for inhibitors of Vif-A3F interactions: SummarysummaryVif-A3F Summary
Description:
The recent discovery and characterization of host restriction factors, which provide a potent defense to invading retroviruses, has provided new targets for antiviral drug development. APOBEC3G (A3G) and APOBEC3F (A3F) are potent cytidine deaminases that block HIV-1 replication; HIV-1 expresses the Vif protein, which counteracts A3G and A3F by binding to them and inducing their proteasomal degradation.

The Vif-A3G and Vif-A3F interactions provide two potential targets for development of small molecules which can interfere with the ability of Vif to degrade A3G or A3F. Structural determinants of these interactions have been mapped to 1 to 5 amino acids in A3G, A3F, and Vif, suggesting that small molecules could potentially inhibit these interactions. To identify inhibitors that can interfere with Vif mediated degradation of A3G and A3F, we have developed Hela cell lines that stably express A3G or A3F tagged with eYFP at the C-terminal end (A3G-eYFP or A3F-eYFP, respectively) and Vif. In the absence of an inhibitor, Vif induces degradation of A3G-eYFP and A3F-eYFP, and the cells are not fluorescent. In the presence of an inhibitor, it is expected that Vif will be unable to degrade A3G-eYFP and/or A3F-eYFP, resulting in an increase in yellow fluorescence. In this assay, Vif A3G was screened against the cherry picks ordered from the MLSMR.

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: MH093245
Assay Submitter (PI): Vinay Pathak, National Cancer Institute
Protocol
The qHTS was performed with Hela_A3G_Vif_eYFP stable cells (A3G-Vif), that showed enhanced yellow fluorescent protein (eYFP) after compound treatment during automated liquid handling in 1,536-well format. A3G-Vif cells were harvested from T225 flask and resuspended in phenol free OPTI-MEM medium with 5% FBS. Then 5 uL of 140,000 cells/mL resuspended cells was dispensed into each well of low base Aurora COC 1,536-well plates (black walled, clear bottom) using a Multidrop Combi dispenser. After overnight culture at 37 deg C with 5% CO2, a total of 23 nL of compounds at 5 selected concentrations from the NPC or positive control (2 mM stock of MG132) in DMSO was transferred to each well of the assay plate using a pintool (Kalypsys, San Diego, CA), and the plates were further incubated at 37 deg C with 5% CO2 for 24 hours. After that, the plates were measured on Acumen Explorer (TTP Lab Tech). The fluorescence was detected by 488 nm/Ex and 500-530nM/Em.
Comment
Compound Ranking:

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.
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Activity_ScoreActivity score.Integer
6Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.00274 uM (0.00273854μM**)% Activity at given concentration.Float%
16Activity at 0.014 uM (0.0136617μM**)% Activity at given concentration.Float%
17Activity at 0.068 uM (0.0684325μM**)% Activity at given concentration.Float%
18Activity at 0.342 uM (0.342472μM**)% Activity at given concentration.Float%
19Activity at 1.707 uM (1.70733μM**)% Activity at given concentration.Float%
20Activity at 8.557 uM (8.55677μM**)% Activity at given concentration.Float%
21Activity at 36.90 uM (36.9μM**)% Activity at given concentration.Float%
22Activity at 46.10 uM (46.1μM**)% Activity at given concentration.Float%
23Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH093245

Data Table (Concise)
Classification
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