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BioAssay: AID 720636

qHTS assay for small molecule agonists of the antioxidant response element (ARE) signaling pathway measured by Nrf2-dependant transcriptional activity

The antioxidant response element (ARE) signaling pathway plays an important role in the cellular anti-oxidant defense. Exposure to stressors which are exogenous like ionizing radiations, chemicals, U-V light, and endogenous like cell signaling, metabolic processes or inflammation, reactive oxygen species (ROS), can induce oxidative stress in the cells. Oxidative stress has been implicated in the pathogenesis of a variety of diseases ranging from cancer to neurodegeneration. ..more
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 Tested Compounds
 Tested Compounds
All(2333)
 
 
Active(44)
 
 
Inactive(2007)
 
 
Inconclusive(349)
 
 
 Tested Substances
 Tested Substances
All(2864)
 
 
Active(49)
 
 
Inactive(2413)
 
 
Inconclusive(402)
 
 
 Related BioAssays
 Related BioAssays
AID: 720636
Data Source: NCGC (NRF205)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2013-09-10

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 44
Description:
National Center for Advancing Translational Sciences [NCATS]
NIH Chemical Genomics Center [NCGC]
U.S. Environmental Protection Agency [EPA]
National Institutes of Environmental Health Sciences [NIEHS]
National Toxicology Program [NTP]

Assay Overview:

The antioxidant response element (ARE) signaling pathway plays an important role in the cellular anti-oxidant defense. Exposure to stressors which are exogenous like ionizing radiations, chemicals, U-V light, and endogenous like cell signaling, metabolic processes or inflammation, reactive oxygen species (ROS), can induce oxidative stress in the cells. Oxidative stress has been implicated in the pathogenesis of a variety of diseases ranging from cancer to neurodegeneration.

ARE-luc HepG2 cell line (Simmons et al., Current Chemical Genomics, 5:1-12, 2011) has been used for identifying the chemicals that induce oxidative stress. The cell line has an Nrf2-responsive luciferase reporter gene that has been engineered to measure Nrf2-dependant transcriptional activity.
Protocol
Assay Protocol Summary:
For measuring ARE induction; 2,000 ARE-luc HepG2 cells in 5 uL of assay medium containing 1% dialyzed FBS per well were dispensed into white wall/solid bottom 1536-well plates using a Flying Reagent Dispenser (FRD) (Aurora Discovery). The assay plates were incubated for 6 hr at 37 C to allow cell attachment to the well bottom, followed by the transfer of 23 nL compound or DMSO vehicle by a pin tool (Kalypsys). The plates were incubated for 16 hr at 37 C and 5% CO2, followed by the addition of 5 uL of ONE-GloTM luciferase reagent (Promega) into each well using FRD. After 30 min room temperature incubation, plates were read on a ViewLux plate reader (PerkinElmer).
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 activators are ranked higher than compounds that showed apparent inhibition.
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.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Cell Type: HEPG2
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Phenotype-Replicate_1Indicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency-Replicate_1*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
3Efficacy-Replicate_1Maximal 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 Comment-Replicate_1Annotation/notes on a particular compound's data or its analysis.String
5Activity_Score-Replicate_1Activity score.Integer
6Curve_Description-Replicate_1A 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_LogAC50-Replicate_1The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlope-Replicate_1The Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2-Replicate_1R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivity-Replicate_1The asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivity-Replicate_1Efficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClass-Replicate_1Numerical encoding of curve description for the fitted Hill equation.Float
13Excluded_Points-Replicate_1Which 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_Response-Replicate_1Maximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0005219194 uM-Replicate_1 (0.000521919μM**)% Activity at given concentration.Float%
16Activity at 0.00155 uM-Replicate_1 (0.00155426μM**)% Activity at given concentration.Float%
17Activity at 0.00461 uM-Replicate_1 (0.00461369μM**)% Activity at given concentration.Float%
18Activity at 0.00590 uM-Replicate_1 (0.005901μM**)% Activity at given concentration.Float%
19Activity at 0.014 uM-Replicate_1 (0.0140515μM**)% Activity at given concentration.Float%
20Activity at 0.041 uM-Replicate_1 (0.0414187μM**)% Activity at given concentration.Float%
21Activity at 0.068 uM-Replicate_1 (0.0684614μM**)% Activity at given concentration.Float%
22Activity at 0.128 uM-Replicate_1 (0.127554μM**)% Activity at given concentration.Float%
23Activity at 0.377 uM-Replicate_1 (0.377223μM**)% Activity at given concentration.Float%
24Activity at 0.782 uM-Replicate_1 (0.781581μM**)% Activity at given concentration.Float%
25Activity at 1.172 uM-Replicate_1 (1.17239μM**)% Activity at given concentration.Float%
26Activity at 3.424 uM-Replicate_1 (3.42413μM**)% Activity at given concentration.Float%
27Activity at 5.843 uM-Replicate_1 (5.84295μM**)% Activity at given concentration.Float%
28Activity at 10.19 uM-Replicate_1 (10.194μM**)% Activity at given concentration.Float%
29Activity at 29.79 uM-Replicate_1 (29.7915μM**)% Activity at given concentration.Float%
30Activity at 43.68 uM-Replicate_1 (43.6836μM**)% Activity at given concentration.Float%
31Activity at 92.36 uM-Replicate_1 (92.3584μM**)% Activity at given concentration.Float%
32Activity at 218.9 uM-Replicate_1 (218.902μM**)% Activity at given concentration.Float%
33Activity at 489.5 uM-Replicate_1 (489.512μM**)% Activity at given concentration.Float%
34Activity at 1094.5 uM-Replicate_1 (1094.48μM**)% Activity at given concentration.Float%
35Activity at 2447.1 uM-Replicate_1 (2447.06μM**)% Activity at given concentration.Float%
36Activity at 5812.0 uM-Replicate_1 (5812μM**)% Activity at given concentration.Float%
37Activity at 13000.0 uM-Replicate_1 (13000μM**)% Activity at given concentration.Float%
38Activity at 29060.0 uM-Replicate_1 (29060μM**)% Activity at given concentration.Float%
39Activity at 64980.0 uM-Replicate_1 (64980μM**)% Activity at given concentration.Float%
40Compound QC-Replicate_1NCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String
41Phenotype-Replicate_2Indicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
42Potency-Replicate_2Concentration 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
43Efficacy-Replicate_2Maximal 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%
44Analysis Comment-Replicate_2Annotation/notes on a particular compound's data or its analysis.String
45Activity_Score-Replicate_2Activity score.Integer
46Curve_Description-Replicate_2A 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
47Fit_LogAC50-Replicate_2The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
48Fit_HillSlope-Replicate_2The Hill slope from a fit of the data to the Hill equation.Float
49Fit_R2-Replicate_2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
50Fit_InfiniteActivity-Replicate_2The asymptotic efficacy from a fit of the data to the Hill equation.Float%
51Fit_ZeroActivity-Replicate_2Efficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
52Fit_CurveClass-Replicate_2Numerical encoding of curve description for the fitted Hill equation.Float
53Excluded_Points-Replicate_2Which 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
54Max_Response-Replicate_2Maximum activity observed for compound (usually at highest concentration tested).Float%
55Activity at 0.0005219194 uM-Replicate_2 (0.000521919μM**)% Activity at given concentration.Float%
56Activity at 0.00155 uM-Replicate_2 (0.00155426μM**)% Activity at given concentration.Float%
57Activity at 0.00461 uM-Replicate_2 (0.00461369μM**)% Activity at given concentration.Float%
58Activity at 0.00590 uM-Replicate_2 (0.005901μM**)% Activity at given concentration.Float%
59Activity at 0.014 uM-Replicate_2 (0.0140515μM**)% Activity at given concentration.Float%
60Activity at 0.041 uM-Replicate_2 (0.0414187μM**)% Activity at given concentration.Float%
61Activity at 0.068 uM-Replicate_2 (0.0684614μM**)% Activity at given concentration.Float%
62Activity at 0.128 uM-Replicate_2 (0.127554μM**)% Activity at given concentration.Float%
63Activity at 0.377 uM-Replicate_2 (0.377223μM**)% Activity at given concentration.Float%
64Activity at 0.782 uM-Replicate_2 (0.781581μM**)% Activity at given concentration.Float%
65Activity at 1.172 uM-Replicate_2 (1.17239μM**)% Activity at given concentration.Float%
66Activity at 3.424 uM-Replicate_2 (3.42413μM**)% Activity at given concentration.Float%
67Activity at 5.843 uM-Replicate_2 (5.84295μM**)% Activity at given concentration.Float%
68Activity at 10.19 uM-Replicate_2 (10.194μM**)% Activity at given concentration.Float%
69Activity at 29.79 uM-Replicate_2 (29.7915μM**)% Activity at given concentration.Float%
70Activity at 43.68 uM-Replicate_2 (43.6836μM**)% Activity at given concentration.Float%
71Activity at 92.36 uM-Replicate_2 (92.3584μM**)% Activity at given concentration.Float%
72Activity at 218.9 uM-Replicate_2 (218.902μM**)% Activity at given concentration.Float%
73Activity at 489.5 uM-Replicate_2 (489.512μM**)% Activity at given concentration.Float%
74Activity at 1094.5 uM-Replicate_2 (1094.48μM**)% Activity at given concentration.Float%
75Activity at 2447.1 uM-Replicate_2 (2447.06μM**)% Activity at given concentration.Float%
76Activity at 5812.0 uM-Replicate_2 (5812μM**)% Activity at given concentration.Float%
77Activity at 13000.0 uM-Replicate_2 (13000μM**)% Activity at given concentration.Float%
78Activity at 29060.0 uM-Replicate_2 (29060μM**)% Activity at given concentration.Float%
79Activity at 64980.0 uM-Replicate_2 (64980μM**)% Activity at given concentration.Float%

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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