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BioAssay: AID 651778

qHTS assay for small molecule agonists of the peroxisome proliferator-activated receptor alpha (PPARalpha) signaling pathway

The Peroxisome proliferator-activated receptors (PPARs) are lipid-activated transcription factors of the nuclear receptor superfamily with three distinct subtypes (alpha, garmma and beta/delta). All these subtypes heterodimerize with retinoid X receptor and have different physiological functions. PPARalpha is highly expressed in tissues with high rates of mitochondrial fatty acid oxidation, such as liver, heart, muscle, kidney and cells of the arterial wall. PPARalpha is a major regulator of lipid metabolism and energy homeostasis. ..more
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 Tested Compounds
 Tested Compounds
All(2329)
 
 
Active(28)
 
 
Inactive(2258)
 
 
Inconclusive(58)
 
 
 Tested Substances
 Tested Substances
All(2864)
 
 
Active(33)
 
 
Inactive(2772)
 
 
Inconclusive(59)
 
 
 Related BioAssays
 Related BioAssays
AID: 651778
Data Source: NCGC (PPARA536)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
Deposit Date: 2012-11-15

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 28
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 Peroxisome proliferator-activated receptors (PPARs) are lipid-activated transcription factors of the nuclear receptor superfamily with three distinct subtypes (alpha, garmma and beta/delta). All these subtypes heterodimerize with retinoid X receptor and have different physiological functions. PPARalpha is highly expressed in tissues with high rates of mitochondrial fatty acid oxidation, such as liver, heart, muscle, kidney and cells of the arterial wall. PPARalpha is a major regulator of lipid metabolism and energy homeostasis.

A cell based assay was used to identify activators of PPARalpha. The assay detects binding of a ligand to PPARalpha leading to its activation, translocation and complementation thus forming a functional enzyme capable of hydrolyzing the substrate molecule and generating chemiluminescent signal.
Protocol
Assay Protocol Summary:
2,000 cells in 6 uL/well were dispensed into white wall/solid bottom 1536-well plates using a Multidrop Combi (Thermo Scientific) dispenser. After incubation 37 C and 5% CO2 overnight, 23 nL compound or DMSO only were transferred by a pin tool. The plates were incubated for 6 hr at 37 C and 5% CO2. 3 ul PathHunter detection reagent (Discoverx) was then added to each well and following 60 min incubation at room temperature, chemiluminescence was quantified on a ViewLux CCD-based 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.
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
5Activity_ScoreActivity score.Integer
6Curve_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
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_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
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0004339321 uM (0.000433932μM**)% Activity at given concentration.Float%
16Activity at 0.00129 uM (0.00129225μM**)% Activity at given concentration.Float%
17Activity at 0.00384 uM (0.0038359μM**)% Activity at given concentration.Float%
18Activity at 0.00491 uM (0.004906μM**)% Activity at given concentration.Float%
19Activity at 0.012 uM (0.0116724μM**)% Activity at given concentration.Float%
20Activity at 0.034 uM (0.0344363μM**)% Activity at given concentration.Float%
21Activity at 0.057 uM (0.0569208μM**)% Activity at given concentration.Float%
22Activity at 0.106 uM (0.106051μM**)% Activity at given concentration.Float%
23Activity at 0.314 uM (0.313627μM**)% Activity at given concentration.Float%
24Activity at 0.650 uM (0.649777μM**)% Activity at given concentration.Float%
25Activity at 0.975 uM (0.974763μM**)% Activity at given concentration.Float%
26Activity at 2.847 uM (2.84689μM**)% Activity at given concentration.Float%
27Activity at 4.858 uM (4.85796μM**)% Activity at given concentration.Float%
28Activity at 8.476 uM (8.47551μM**)% Activity at given concentration.Float%
29Activity at 24.77 uM (24.7692μM**)% Activity at given concentration.Float%
30Activity at 36.32 uM (36.3186μM**)% Activity at given concentration.Float%
31Activity at 76.79 uM (76.7883μM**)% Activity at given concentration.Float%
32Activity at 182.0 uM (182.021μM**)% Activity at given concentration.Float%
33Activity at 407.0 uM (407.01μM**)% Activity at given concentration.Float%
34Activity at 910.0 uM (909.998μM**)% Activity at given concentration.Float%
35Activity at 2034.8 uM (2034.82μM**)% Activity at given concentration.Float%
36Activity at 4832.0 uM (4832μM**)% Activity at given concentration.Float%
37Activity at 10810.0 uM (10810μM**)% Activity at given concentration.Float%
38Activity at 24160.0 uM (24160μM**)% Activity at given concentration.Float%
39Activity at 54020.0 uM (54020μM**)% Activity at given concentration.Float%
40Compound 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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