Bookmark and Share
BioAssay: AID 651743

qHTS assay for small molecule activators of the p53 signaling pathway

p53, a tumor suppressor protein, is activated following cellular insult, including DNA damage and other cellular stresses. The activation of p53 regulates cell fate by inducing DNA repair, cell cycle arrest, apoptosis, or cellular senescence. The activation of p53, therefore, is a good indicator of DNA damage and other cellular stresses. The CellSensor p53RE-bla HCT-116 cell line (Invitrogen) more ..
_
   
 Tested Compounds
 Tested Compounds
All(2329)
 
 
Active(73)
 
 
Inactive(2084)
 
 
Inconclusive(224)
 
 
 Tested Substances
 Tested Substances
All(2864)
 
 
Active(88)
 
 
Inactive(2530)
 
 
Inconclusive(246)
 
 
 Related BioAssays
 Related BioAssays
AID: 651743
Data Source: NCGC (P53298)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2012-11-01

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 73
Depositor Specified Assays
AIDNameTypeComment
743292qHTS assay for cell viability of the p53RE-bla HCT-116 cell lineconfirmatory
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:

p53, a tumor suppressor protein, is activated following cellular insult, including DNA damage and other cellular stresses. The activation of p53 regulates cell fate by inducing DNA repair, cell cycle arrest, apoptosis, or cellular senescence. The activation of p53, therefore, is a good indicator of DNA damage and other cellular stresses. The CellSensor p53RE-bla HCT-116 cell line (Invitrogen) contains a stably integrated beta-lactamase reporter gene under control of the p53 response elements. The activation of the report gene under culture conditions can be detected by fluorescence intensity. This cell line can be used to screen chemicals in a 1536-well plate format and to identify agonists that activate p53 signaling pathway.
Protocol
Assay Protocol Summary:

The p53RE-bla cells were dispensed at 4,000 cells/5 ul/well in 1536-well black wall/clear bottom plates using a Multidrop dispenser. After the assay plates were incubated at a 37 C/5% CO2 incubator for 5 hours, 23 nL of compounds dissolved in DMSO, positive controls or DMSO only was transferred to the assay plate by a pin tool. The plates were incubated at 37 C for 16 hours. After 1 uL of LiveBLAzer B/G FRET substrate was added using a Flying Reagent Dispenser, the plates were incubated at room temperature for 2 hours, and fluorescence intensity was measured by an Envision plate reader.
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.
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
5W460-Activity_ScoreActivity score.Integer
6W460-Curve_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
7W460-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8W460-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9W460-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10W460-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11W460-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12W460-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13W460-Excluded_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
14W460-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15W460-Activity at 0.0005213369 uM (0.000521337μM**)% Activity at given concentration.Float%
16W460-Activity at 0.00156 uM (0.0015582μM**)% Activity at given concentration.Float%
17W460-Activity at 0.00466 uM (0.0046561μM**)% Activity at given concentration.Float%
18W460-Activity at 0.00590 uM (0.00590092μM**)% Activity at given concentration.Float%
19W460-Activity at 0.014 uM (0.0140534μM**)% Activity at given concentration.Float%
20W460-Activity at 0.042 uM (0.0417584μM**)% Activity at given concentration.Float%
21W460-Activity at 0.068 uM (0.0684683μM**)% Activity at given concentration.Float%
22W460-Activity at 0.127 uM (0.126926μM**)% Activity at given concentration.Float%
23W460-Activity at 0.378 uM (0.378214μM**)% Activity at given concentration.Float%
24W460-Activity at 0.782 uM (0.781568μM**)% Activity at given concentration.Float%
25W460-Activity at 1.156 uM (1.15581μM**)% Activity at given concentration.Float%
26W460-Activity at 3.420 uM (3.41952μM**)% Activity at given concentration.Float%
27W460-Activity at 5.843 uM (5.84314μM**)% Activity at given concentration.Float%
28W460-Activity at 10.21 uM (10.2119μM**)% Activity at given concentration.Float%
29W460-Activity at 30.24 uM (30.2439μM**)% Activity at given concentration.Float%
30W460-Activity at 43.68 uM (43.6843μM**)% Activity at given concentration.Float%
31W460-Activity at 92.27 uM (92.2654μM**)% Activity at given concentration.Float%
32W460-Activity at 218.9 uM (218.915μM**)% Activity at given concentration.Float%
33W460-Activity at 489.5 uM (489.494μM**)% Activity at given concentration.Float%
34W460-Activity at 1094.5 uM (1094.51μM**)% Activity at given concentration.Float%
35W460-Activity at 2447.3 uM (2447.32μM**)% Activity at given concentration.Float%
36W460-Activity at 5812.2 uM (5812.25μM**)% Activity at given concentration.Float%
37W460-Activity at 12996.2 uM (12996.2μM**)% Activity at given concentration.Float%
38W460-Activity at 29059.5 uM (29059.5μM**)% Activity at given concentration.Float%
39W460-Activity at 64977.0 uM (64977μM**)% Activity at given concentration.Float%
40Ratio-Activity_ScoreActivity score.Integer
41Ratio-Curve_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
42Ratio-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
43Ratio-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
44Ratio-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
45Ratio-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
46Ratio-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
47Ratio-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
48Ratio-Excluded_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
49Ratio-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
50Ratio-Activity at 0.0005213369 uM (0.000521337μM**)% Activity at given concentration.Float%
51Ratio-Activity at 0.00156 uM (0.0015582μM**)% Activity at given concentration.Float%
52Ratio-Activity at 0.00466 uM (0.0046561μM**)% Activity at given concentration.Float%
53Ratio-Activity at 0.00590 uM (0.00590092μM**)% Activity at given concentration.Float%
54Ratio-Activity at 0.014 uM (0.0140534μM**)% Activity at given concentration.Float%
55Ratio-Activity at 0.042 uM (0.0417584μM**)% Activity at given concentration.Float%
56Ratio-Activity at 0.068 uM (0.0684683μM**)% Activity at given concentration.Float%
57Ratio-Activity at 0.127 uM (0.126926μM**)% Activity at given concentration.Float%
58Ratio-Activity at 0.378 uM (0.378214μM**)% Activity at given concentration.Float%
59Ratio-Activity at 0.782 uM (0.781568μM**)% Activity at given concentration.Float%
60Ratio-Activity at 1.156 uM (1.15581μM**)% Activity at given concentration.Float%
61Ratio-Activity at 3.420 uM (3.41952μM**)% Activity at given concentration.Float%
62Ratio-Activity at 5.843 uM (5.84314μM**)% Activity at given concentration.Float%
63Ratio-Activity at 10.21 uM (10.2119μM**)% Activity at given concentration.Float%
64Ratio-Activity at 30.24 uM (30.2439μM**)% Activity at given concentration.Float%
65Ratio-Activity at 43.68 uM (43.6843μM**)% Activity at given concentration.Float%
66Ratio-Activity at 92.27 uM (92.2654μM**)% Activity at given concentration.Float%
67Ratio-Activity at 218.9 uM (218.915μM**)% Activity at given concentration.Float%
68Ratio-Activity at 489.5 uM (489.494μM**)% Activity at given concentration.Float%
69Ratio-Activity at 1094.5 uM (1094.51μM**)% Activity at given concentration.Float%
70Ratio-Activity at 2447.3 uM (2447.32μM**)% Activity at given concentration.Float%
71Ratio-Activity at 5812.2 uM (5812.25μM**)% Activity at given concentration.Float%
72Ratio-Activity at 12996.2 uM (12996.2μM**)% Activity at given concentration.Float%
73Ratio-Activity at 29059.5 uM (29059.5μM**)% Activity at given concentration.Float%
74Ratio-Activity at 64977.0 uM (64977μM**)% Activity at given concentration.Float%
75W530-Activity_ScoreActivity score.Integer
76W530-Curve_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
77W530-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
78W530-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
79W530-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
80W530-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
81W530-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
82W530-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
83W530-Excluded_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
84W530-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
85W530-Activity at 0.0005213369 uM (0.000521337μM**)% Activity at given concentration.Float%
86W530-Activity at 0.00156 uM (0.0015582μM**)% Activity at given concentration.Float%
87W530-Activity at 0.00466 uM (0.0046561μM**)% Activity at given concentration.Float%
88W530-Activity at 0.00590 uM (0.00590092μM**)% Activity at given concentration.Float%
89W530-Activity at 0.014 uM (0.0140534μM**)% Activity at given concentration.Float%
90W530-Activity at 0.042 uM (0.0417584μM**)% Activity at given concentration.Float%
91W530-Activity at 0.068 uM (0.0684683μM**)% Activity at given concentration.Float%
92W530-Activity at 0.127 uM (0.126926μM**)% Activity at given concentration.Float%
93W530-Activity at 0.378 uM (0.378214μM**)% Activity at given concentration.Float%
94W530-Activity at 0.782 uM (0.781568μM**)% Activity at given concentration.Float%
95W530-Activity at 1.156 uM (1.15581μM**)% Activity at given concentration.Float%
96W530-Activity at 3.420 uM (3.41952μM**)% Activity at given concentration.Float%
97W530-Activity at 5.843 uM (5.84314μM**)% Activity at given concentration.Float%
98W530-Activity at 10.21 uM (10.2119μM**)% Activity at given concentration.Float%
99W530-Activity at 30.24 uM (30.2439μM**)% Activity at given concentration.Float%
100W530-Activity at 43.68 uM (43.6843μM**)% Activity at given concentration.Float%
101W530-Activity at 92.27 uM (92.2654μM**)% Activity at given concentration.Float%
102W530-Activity at 218.9 uM (218.915μM**)% Activity at given concentration.Float%
103W530-Activity at 489.5 uM (489.494μM**)% Activity at given concentration.Float%
104W530-Activity at 1094.5 uM (1094.51μM**)% Activity at given concentration.Float%
105W530-Activity at 2447.3 uM (2447.32μM**)% Activity at given concentration.Float%
106W530-Activity at 5812.2 uM (5812.25μM**)% Activity at given concentration.Float%
107W530-Activity at 12996.2 uM (12996.2μM**)% Activity at given concentration.Float%
108W530-Activity at 29059.5 uM (29059.5μM**)% Activity at given concentration.Float%
109W530-Activity at 64977.0 uM (64977μM**)% Activity at given concentration.Float%
110Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
Classification
PageFrom: