Bookmark and Share
BioAssay: AID 602310

qHTS for Inhibitors of Vif-A3G Interactions: qHTS

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
_
   
 Tested Compounds
 Tested Compounds
All(402340)
 
 
Active(311)
 
 
Inactive(393873)
 
 
Inconclusive(8244)
 
 
 Tested Substances
 Tested Substances
All(406764)
 
 
Active(311)
 
 
Inactive(398161)
 
 
Inconclusive(8292)
 
 
AID: 602310
Data Source: NCGC (VIFG100)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2012-03-08
Modify Date: 2012-11-27

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 311
Depositor Specified Assays
AIDNameTypeComment
588444qHTS for inhibitors of Vif-A3G interactions: SummarysummarySummary AID
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 assays Vif A3G was screened against the Molecular Library Small Molecule Repository (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
5 uL of 700 cells/well of A3G-YFP cells are dispensed into black, clear bottom plates. The plates are incubated overnight at 37 deg C. 23 nL of compounds are added by pintool. The plates are incubated overnight at 37 deg C and then read on the TTP Acumen (Ex 488/ Em 500-530).
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
Show more
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
5Curve_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
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_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
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.092 uM (0.0922μM**)% Activity at given concentration.Float%
15Activity at 0.184 uM (0.184μM**)% Activity at given concentration.Float%
16Activity at 0.369 uM (0.369279μM**)% Activity at given concentration.Float%
17Activity at 0.737 uM (0.737μM**)% Activity at given concentration.Float%
18Activity at 0.922 uM (0.922μM**)% Activity at given concentration.Float%
19Activity at 1.840 uM (1.84μM**)% Activity at given concentration.Float%
20Activity at 2.300 uM (2.3μM**)% Activity at given concentration.Float%
21Activity at 3.690 uM (3.69μM**)% Activity at given concentration.Float%
22Activity at 4.610 uM (4.61μM**)% Activity at given concentration.Float%
23Activity at 9.220 uM (9.22μM**)% Activity at given concentration.Float%
24Activity at 11.50 uM (11.5μM**)% Activity at given concentration.Float%
25Activity at 19.82 uM (19.8208μM**)% Activity at given concentration.Float%
26Activity at 46.10 uM (46.1μM**)% Activity at given concentration.Float%
27Activity at 92.20 uM (92.2μM**)% Activity at given concentration.Float%
28Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

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

Data Table (Concise)
Classification
PageFrom: