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BioAssay: AID 915

qHTS Assay for Identification of Small Molecule Antagonists for Hypoxia Response Element Signaling Pathway

Hypoxia-inducible factor-1 (HIF-1) is the major hypoxia-regulated transcription factor that controls cellular responses to low oxygen concentration. HIF-1 is composed of two subunits: hypoxia responsive HIF-1 alpha and constitutively expressed HIF-1 beta, which is also known as aryl hydrocarbon receptor nuclear translocator. During hypoxic conditions, HIF-1 alpha heterodimerizes with HIF-1 beta more ..
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 Tested Compounds
 Tested Compounds
All(10838)
 
 
Active(442)
 
 
Inactive(7611)
 
 
Inconclusive(2954)
 
 
 Tested Substances
 Tested Substances
All(11410)
 
 
Active(456)
 
 
Inactive(7914)
 
 
Inconclusive(3040)
 
 
 Related BioAssays
 Related BioAssays
AID: 915
Data Source: NCGC (HRER229i)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Screening Center Network
BioAssay Version:
Deposit Date: 2007-12-19
Modify Date: 2010-07-06

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 442
Related Experiments
AIDNameTypeComment
914qHTS Assay for Identification of Small Molecule Agonists for Hypoxia Response Element Signaling PathwayConfirmatorydepositor-specified cross reference
2120qHTS Assay for Identification of Small Molecule Inducers of Hypoxia ResponseConfirmatorydepositor-specified cross reference
Description:
NIH Molecular Libraries Screening Centers Network [MLSCN]
NIH Chemical Genomics Center [NCGC]

MLSCN Grant: None

NCGC Assay Overview:

Hypoxia-inducible factor-1 (HIF-1) is the major hypoxia-regulated transcription factor that controls cellular responses to low oxygen concentration. HIF-1 is composed of two subunits: hypoxia responsive HIF-1 alpha and constitutively expressed HIF-1 beta, which is also known as aryl hydrocarbon receptor nuclear translocator. During hypoxic conditions, HIF-1 alpha heterodimerizes with HIF-1 beta and translocates into the nucleus where the HIF-1 complex binds to the hypoxia-response element (HRE) and activates expression of target genes such as VEGF (vascular endothelial growth factor). HIF-1 alpha protein levels are elevated in many solid tumors, including those of the cervix and brain. Within tumors, cells that are the greatest distance from blood vessels and therefore the most hypoxic express the highest levels of HIF-1 alpha. Therapeutic blockade of the HIF-1 signaling pathway in cancer cells therefore provides an attractive strategy for development of anticancer drugs.

Using a beta-lactamase reporter-gene under control of the hypoxia response element, a cell-based assay [CellSensor(TM) HRE-bla ME-180 developed by Invitrogen Corp.] was used to measure hypoxia-induced signaling. Library compounds were measured for their ability to negatively modulate the action of EC50 CoCl2 on reporter gene activity. Compounds were screened in a titration series in 1536-well format. Negative modulators were identified based on the AC50 values derived from the screening data.
Protocol
NCGC Assay Protocol Summary:
2500 cells/5uL in Opti-MEM medium containing 0.5% dialyzed FBS, 0.1 mM NEAA, 1 mM sodium pyruvate and 10 mM HEPES was dispensed into 1536-well plates and cells were cultured at 37 degrees Celsius overnight. Next day 23 nL of compounds or DMSO were delivered to each well using a pin tool, and 1uL of EC50 level of CoCl2, or EC100 level of CoCl2, or assay medium was added. And then the plates were incubated at 37 degrees Celsius for 17 hours. 1 uL of CCF4-dye mixture was added to each well. After incubated at room temperature for 2.5 hours the plates were measured on an EnVision plate reader at Excitation 405nm, Emission1=460nm and Emission 2=530nm. The %Activity was determined from the ratio of 460nm/530nm.
Comment
Compound Ranking:
1. Compounds are first classified as being probable inhibitors (compounds decreasing signal), fluorescent (artifactual results), cytotoxic (artifactual results), inactive, or inconclusive based on the ratio and component channel readouts.
2. Compounds with the inhibitor phenotype were ranked by efficacy and potency from 99 to 20. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Cell Type: ME-180
From ChEMBL:
Assay Type: Functional
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
5Ratio-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 signficant, but below 80% of control.String
6Ratio-Fit_LogAC50The logarithm of the AC50 (calculated based on Molar units) from a fit of the ratio data to the Hill equation.Float
7Ratio-Fit_HillSlopeThe Hill slope from a fit of the ratio data to the Hill equation.Float
8Ratio-Fit_R2R^2 fit value of ratio curve. Closer to 1.0 equates to better Hill equation fit.Float
9Ratio-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the ratio data to the Hill equation.Float%
10Ratio-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the ratio data to the Hill equation.Float%
11Ratio-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Ratio-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
13Ratio-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Ratio-Activity at 0.0005000000 uM (0.0005μM**)% Activity at given concentration.Float%
15Ratio-Activity at 0.00110 uM (0.0011μM**)% Activity at given concentration.Float%
16Ratio-Activity at 0.00250 uM (0.0025μM**)% Activity at given concentration.Float%
17Ratio-Activity at 0.00550 uM (0.0055μM**)% Activity at given concentration.Float%
18Ratio-Activity at 0.012 uM (0.0123μM**)% Activity at given concentration.Float%
19Ratio-Activity at 0.028 uM (0.0275μM**)% Activity at given concentration.Float%
20Ratio-Activity at 0.062 uM (0.0616μM**)% Activity at given concentration.Float%
21Ratio-Activity at 0.138 uM (0.1376μM**)% Activity at given concentration.Float%
22Ratio-Activity at 0.308 uM (0.3077μM**)% Activity at given concentration.Float%
23Ratio-Activity at 0.688 uM (0.6881μM**)% Activity at given concentration.Float%
24Ratio-Activity at 1.539 uM (1.5386μM**)% Activity at given concentration.Float%
25Ratio-Activity at 3.440 uM (3.4404μM**)% Activity at given concentration.Float%
26Ratio-Activity at 7.693 uM (7.6928μM**)% Activity at given concentration.Float%
27Ratio-Activity at 17.20 uM (17.201μM**)% Activity at given concentration.Float%
28Ratio-Activity at 38.46 uM (38.4615μM**)% Activity at given concentration.Float%
29W460-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 signficant, but below 80% of control.String
30W460-Fit_LogAC50The logarithm of the AC50 (calculated based on Molar units) from a fit of the ratio data to the Hill equation.Float
31W460-Fit_HillSlopeThe Hill slope from a fit of the ratio data to the Hill equation.Float
32W460-Fit_R2R^2 fit value of ratio curve. Closer to 1.0 equates to better Hill equation fit.Float
33W460-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the ratio data to the Hill equation.Float%
34W460-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the ratio data to the Hill equation.Float%
35W460-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
36W460-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
37W460-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
38W460-Activity at 0.0005000000 uM (0.0005μM**)% Activity at given concentration.Float%
39W460-Activity at 0.00110 uM (0.0011μM**)% Activity at given concentration.Float%
40W460-Activity at 0.00250 uM (0.0025μM**)% Activity at given concentration.Float%
41W460-Activity at 0.00550 uM (0.0055μM**)% Activity at given concentration.Float%
42W460-Activity at 0.012 uM (0.0123μM**)% Activity at given concentration.Float%
43W460-Activity at 0.028 uM (0.0275μM**)% Activity at given concentration.Float%
44W460-Activity at 0.062 uM (0.0616μM**)% Activity at given concentration.Float%
45W460-Activity at 0.138 uM (0.1376μM**)% Activity at given concentration.Float%
46W460-Activity at 0.308 uM (0.3077μM**)% Activity at given concentration.Float%
47W460-Activity at 0.688 uM (0.6881μM**)% Activity at given concentration.Float%
48W460-Activity at 1.539 uM (1.5386μM**)% Activity at given concentration.Float%
49W460-Activity at 3.440 uM (3.4404μM**)% Activity at given concentration.Float%
50W460-Activity at 7.693 uM (7.6928μM**)% Activity at given concentration.Float%
51W460-Activity at 17.20 uM (17.201μM**)% Activity at given concentration.Float%
52W460-Activity at 38.46 uM (38.4615μM**)% Activity at given concentration.Float%
53W530-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 signficant, but below 80% of control.String
54W530-Fit_LogAC50The logarithm of the AC50 (calculated based on Molar units) from a fit of the ratio data to the Hill equation.Float
55W530-Fit_HillSlopeThe Hill slope from a fit of the ratio data to the Hill equation.Float
56W530-Fit_R2R^2 fit value of ratio curve. Closer to 1.0 equates to better Hill equation fit.Float
57W530-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the ratio data to the Hill equation.Float%
58W530-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the ratio data to the Hill equation.Float%
59W530-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
60W530-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
61W530-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
62W530-Activity at 0.0005000000 uM (0.0005μM**)% Activity at given concentration.Float%
63W530-Activity at 0.00110 uM (0.0011μM**)% Activity at given concentration.Float%
64W530-Activity at 0.00250 uM (0.0025μM**)% Activity at given concentration.Float%
65W530-Activity at 0.00550 uM (0.0055μM**)% Activity at given concentration.Float%
66W530-Activity at 0.012 uM (0.0123μM**)% Activity at given concentration.Float%
67W530-Activity at 0.028 uM (0.0275μM**)% Activity at given concentration.Float%
68W530-Activity at 0.062 uM (0.0616μM**)% Activity at given concentration.Float%
69W530-Activity at 0.138 uM (0.1376μM**)% Activity at given concentration.Float%
70W530-Activity at 0.308 uM (0.3077μM**)% Activity at given concentration.Float%
71W530-Activity at 0.688 uM (0.6881μM**)% Activity at given concentration.Float%
72W530-Activity at 1.539 uM (1.5386μM**)% Activity at given concentration.Float%
73W530-Activity at 3.440 uM (3.4404μM**)% Activity at given concentration.Float%
74W530-Activity at 7.693 uM (7.6928μM**)% Activity at given concentration.Float%
75W530-Activity at 17.20 uM (17.201μM**)% Activity at given concentration.Float%
76W530-Activity at 38.46 uM (38.4615μM**)% Activity at given concentration.Float%
77Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

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