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BioAssay: AID 588527

qHTS assay for small molecule agonists of farnesoid X receptor signaling

The farnesoid-X-receptor (FXR), a nuclear hormone receptor is highly expressed in liver, intestine, kidney and adrenal cortex. Natural ligands of FXR are the bile acids (Cholic acid, Chenodeoxy cholic acid etc). It is an important regulator of diverse metabolic pathways, including the bile acid homeostasis, lipid and glucose metabolisms. FXR regulates the expression of target genes by binding more ..
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 Tested Compounds
 Tested Compounds
All(2325)
 
 
Active(3)
 
 
Inactive(2105)
 
 
Inconclusive(313)
 
 
 Tested Substances
 Tested Substances
All(2858)
 
 
Active(3)
 
 
Inactive(2526)
 
 
Inconclusive(329)
 
 
 Related BioAssays
 Related BioAssays
AID: 588527
Data Source: NCGC (FXRA10)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2011-10-15

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 3
Description:
NIH Chemical Genomics Center [NCGC]
U.S. Environmental Protection Agency [EPA]
National Institutes of Environmental Health Sciences [NIEHS]
National Toxicology Program [NTP]

NCGC Assay Overview

The farnesoid-X-receptor (FXR), a nuclear hormone receptor is highly expressed in liver, intestine, kidney and adrenal cortex. Natural ligands of FXR are the bile acids (Cholic acid, Chenodeoxy cholic acid etc). It is an important regulator of diverse metabolic pathways, including the bile acid homeostasis, lipid and glucose metabolisms. FXR regulates the expression of target genes by binding either as a monomer or as a heterodimer with RXR (retinoid X receptor). As FXR regulates a large number of target genes involved in bile acid, lipoprotein and glucose metabolisms, it is a potential pharmacological target for bile acid and lipid disorders. Thus screening environmental chemicals on FXR signaling can be used for the studies focused on metabolic syndrome, hepatic and intestinal toxicity etc.
Protocol
NCGC Assay Protocol Summary
Using a beta-lactamase reporter-gene under control of Farnesoid X receptor (FXR) response element, a cell-based assay [GeneBLAzer FXR-UAS-bla assay developed by Invitrogen] was used to measure FXR ligands signaling. GeneBLAzer FXR-UAS-bla HEK293T cells were used with assay medium containing 10% dialyzed FBS. The assay was performed in clear bottom black Greiner 1536-well plates. Chenodeoxycholic acid and Guggulsterone were used in the screen as agonist and antagonist positive controls respectively. Library compounds were measured for their ability to stimulate the reporter gene activity. Compounds were screened in a titration series in 1536-well format. The fluorescence intensity (405 nm excitation, 460/530 nm emission) was measured using an EnVision plate reader. Data was normalized relative to Chenodeoxycholic acid control (100 uM, 100%), and DMSO only wells (basal, 0%). Concentration-response titration points for each compound were fitted to the Hill equation yielding concentrations of half-maximal stimulation (EC50) and maximal response (efficacy) values.
qHTS protocol for FXR beta-lactamase assay in agonist mode
[Step] [Parameter] [Value] [Description]
1. Reagent; 6 uL; 5000 cells/well
2. Incubation; 37 C for 5-7 hrs
3. Compounds; 23 nL; 0.59 nM to 92 uM
4. Controls (Chenodeoxycholic acid); 23 nL,
- One column with dose response, start 200 uM, 1:2 dilutions in duplicates
- Final concentrations of 100 uM and 60 uM in two columns
5. Incubation; 37 C for 16 hrs
6. Detection reagent; 1 uL of CCF4-dye
7. Incubation; RT for 2 hrs
8. Detection; Fluorescence; 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, active compounds are expected to have the activator phenotype and 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 ChEMBL:
Assay Type: Functional
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
5W460-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
6W460-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7W460-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8W460-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9W460-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10W460-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11W460-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12W460-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
13W460-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14W460-Activity at 0.0004334456 uM (0.000433446μM**)% Activity at given concentration.Float%
15W460-Activity at 0.00130 uM (0.00129551μM**)% Activity at given concentration.Float%
16W460-Activity at 0.00387 uM (0.00387116μM**)% Activity at given concentration.Float%
17W460-Activity at 0.00491 uM (0.00490613μM**)% Activity at given concentration.Float%
18W460-Activity at 0.012 uM (0.0116842μM**)% Activity at given concentration.Float%
19W460-Activity at 0.035 uM (0.0347186μM**)% Activity at given concentration.Float%
20W460-Activity at 0.057 uM (0.0569211μM**)% Activity at given concentration.Float%
21W460-Activity at 0.106 uM (0.105529μM**)% Activity at given concentration.Float%
22W460-Activity at 0.314 uM (0.314453μM**)% Activity at given concentration.Float%
23W460-Activity at 0.650 uM (0.649798μM**)% Activity at given concentration.Float%
24W460-Activity at 0.961 uM (0.960962μM**)% Activity at given concentration.Float%
25W460-Activity at 2.843 uM (2.84305μM**)% Activity at given concentration.Float%
26W460-Activity at 4.858 uM (4.85794μM**)% Activity at given concentration.Float%
27W460-Activity at 8.490 uM (8.49033μM**)% Activity at given concentration.Float%
28W460-Activity at 25.15 uM (25.1453μM**)% Activity at given concentration.Float%
29W460-Activity at 36.32 uM (36.3199μM**)% Activity at given concentration.Float%
30W460-Activity at 76.71 uM (76.7111μM**)% Activity at given concentration.Float%
31W460-Activity at 182.0 uM (182.01μM**)% Activity at given concentration.Float%
32W460-Activity at 407.0 uM (406.974μM**)% Activity at given concentration.Float%
33W460-Activity at 910.0 uM (909.994μM**)% Activity at given concentration.Float%
34W460-Activity at 2034.7 uM (2034.75μM**)% Activity at given concentration.Float%
35W460-Activity at 4832.4 uM (4832.4μM**)% Activity at given concentration.Float%
36W460-Activity at 10805.3 uM (10805.3μM**)% Activity at given concentration.Float%
37W460-Activity at 24160.5 uM (24160.5μM**)% Activity at given concentration.Float%
38W460-Activity at 54023.0 uM (54023μM**)% Activity at given concentration.Float%
39Ratio-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
40Ratio-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
41Ratio-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
42Ratio-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
43Ratio-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
44Ratio-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
45Ratio-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
46Ratio-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
47Ratio-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
48Ratio-Activity at 0.0004334456 uM (0.000433446μM**)% Activity at given concentration.Float%
49Ratio-Activity at 0.00130 uM (0.00129551μM**)% Activity at given concentration.Float%
50Ratio-Activity at 0.00387 uM (0.00387116μM**)% Activity at given concentration.Float%
51Ratio-Activity at 0.00491 uM (0.00490613μM**)% Activity at given concentration.Float%
52Ratio-Activity at 0.012 uM (0.0116842μM**)% Activity at given concentration.Float%
53Ratio-Activity at 0.035 uM (0.0347186μM**)% Activity at given concentration.Float%
54Ratio-Activity at 0.057 uM (0.0569211μM**)% Activity at given concentration.Float%
55Ratio-Activity at 0.106 uM (0.105529μM**)% Activity at given concentration.Float%
56Ratio-Activity at 0.314 uM (0.314453μM**)% Activity at given concentration.Float%
57Ratio-Activity at 0.650 uM (0.649798μM**)% Activity at given concentration.Float%
58Ratio-Activity at 0.961 uM (0.960962μM**)% Activity at given concentration.Float%
59Ratio-Activity at 2.843 uM (2.84305μM**)% Activity at given concentration.Float%
60Ratio-Activity at 4.858 uM (4.85794μM**)% Activity at given concentration.Float%
61Ratio-Activity at 8.490 uM (8.49033μM**)% Activity at given concentration.Float%
62Ratio-Activity at 25.15 uM (25.1453μM**)% Activity at given concentration.Float%
63Ratio-Activity at 36.32 uM (36.3199μM**)% Activity at given concentration.Float%
64Ratio-Activity at 76.71 uM (76.7111μM**)% Activity at given concentration.Float%
65Ratio-Activity at 182.0 uM (182.01μM**)% Activity at given concentration.Float%
66Ratio-Activity at 407.0 uM (406.974μM**)% Activity at given concentration.Float%
67Ratio-Activity at 910.0 uM (909.994μM**)% Activity at given concentration.Float%
68Ratio-Activity at 2034.7 uM (2034.75μM**)% Activity at given concentration.Float%
69Ratio-Activity at 4832.4 uM (4832.4μM**)% Activity at given concentration.Float%
70Ratio-Activity at 10805.3 uM (10805.3μM**)% Activity at given concentration.Float%
71Ratio-Activity at 24160.5 uM (24160.5μM**)% Activity at given concentration.Float%
72Ratio-Activity at 54023.0 uM (54023μM**)% Activity at given concentration.Float%
73W530-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
74W530-Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
75W530-Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
76W530-Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
77W530-Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
78W530-Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
79W530-Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
80W530-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
81W530-Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
82W530-Activity at 0.0004334456 uM (0.000433446μM**)% Activity at given concentration.Float%
83W530-Activity at 0.00130 uM (0.00129551μM**)% Activity at given concentration.Float%
84W530-Activity at 0.00387 uM (0.00387116μM**)% Activity at given concentration.Float%
85W530-Activity at 0.00491 uM (0.00490613μM**)% Activity at given concentration.Float%
86W530-Activity at 0.012 uM (0.0116842μM**)% Activity at given concentration.Float%
87W530-Activity at 0.035 uM (0.0347186μM**)% Activity at given concentration.Float%
88W530-Activity at 0.057 uM (0.0569211μM**)% Activity at given concentration.Float%
89W530-Activity at 0.106 uM (0.105529μM**)% Activity at given concentration.Float%
90W530-Activity at 0.314 uM (0.314453μM**)% Activity at given concentration.Float%
91W530-Activity at 0.650 uM (0.649798μM**)% Activity at given concentration.Float%
92W530-Activity at 0.961 uM (0.960962μM**)% Activity at given concentration.Float%
93W530-Activity at 2.843 uM (2.84305μM**)% Activity at given concentration.Float%
94W530-Activity at 4.858 uM (4.85794μM**)% Activity at given concentration.Float%
95W530-Activity at 8.490 uM (8.49033μM**)% Activity at given concentration.Float%
96W530-Activity at 25.15 uM (25.1453μM**)% Activity at given concentration.Float%
97W530-Activity at 36.32 uM (36.3199μM**)% Activity at given concentration.Float%
98W530-Activity at 76.71 uM (76.7111μM**)% Activity at given concentration.Float%
99W530-Activity at 182.0 uM (182.01μM**)% Activity at given concentration.Float%
100W530-Activity at 407.0 uM (406.974μM**)% Activity at given concentration.Float%
101W530-Activity at 910.0 uM (909.994μM**)% Activity at given concentration.Float%
102W530-Activity at 2034.7 uM (2034.75μM**)% Activity at given concentration.Float%
103W530-Activity at 4832.4 uM (4832.4μM**)% Activity at given concentration.Float%
104W530-Activity at 10805.3 uM (10805.3μM**)% Activity at given concentration.Float%
105W530-Activity at 24160.5 uM (24160.5μM**)% Activity at given concentration.Float%
106W530-Activity at 54023.0 uM (54023μM**)% Activity at given concentration.Float%
107Compound 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
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