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BioAssay: AID 504845

Inhibitors of Regulator of G Protein Signaling (RGS) 4: qHTS

G-protein-coupled receptors (GPCRs) are well-established targets for drug discovery and development. Members of the "regulator of G-protein signaling" (RGS)-protein superfamily have recently emerged as critical modulators of specific GPCR signaling pathways. Via their conserved RGS domain that possesses "GTPase-accelerating protein" (GAP) activity, RGS proteins deactivate heterotrimeric G-protein more ..
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 Tested Compounds
 Tested Compounds
All(385508)
 
 
Active(152)
 
 
Inactive(372601)
 
 
Inconclusive(13170)
 
 
 Tested Substances
 Tested Substances
All(390220)
 
 
Active(152)
 
 
Inactive(376844)
 
 
Inconclusive(13224)
 
 
AID: 504845
Data Source: NCGC (RGS4001)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-06-23

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 152
Related Experiments
AIDNameTypeComment
504856Inhibitors of Regulator of G Protein Signaling (RGS) 4: SummarySummarydepositor-specified cross reference
602388qHTS for Inhibitors of Regulator of G Protein Signaling(RGS) 4: Hit ValidationConfirmatorysame project related to Summary assay
Description:
G-protein-coupled receptors (GPCRs) are well-established targets for drug discovery and development. Members of the "regulator of G-protein signaling" (RGS)-protein superfamily have recently emerged as critical modulators of specific GPCR signaling pathways. Via their conserved RGS domain that possesses "GTPase-accelerating protein" (GAP) activity, RGS proteins deactivate heterotrimeric G-protein alpha (G) subunits and thus attenuate GPCR signal transduction. Combining specific RGS domain inhibitors with existing GPCR agonists should potentiate cellular responses and could also increase the specificity of action of existing therapeutics. Small molecule modulators of RGS domain activity should also have clinical utility in inhibiting or potentiating the actions of endogenous GPCR agonists. In particular, the diversity of RGS proteins with highly localized and dynamically regulated distributions in the human brain makes this family of proteins attractive targets for pharmacotherapy of central nervous system (CNS) disorders such as Parkinson's disease, schizophrenia, and anxiety.
Hence, in a collaboration between the University of North Caroline Chapel Hill (School of Medicine) and NIH Chemical Genomics Center (NCGC) a novel HTS-compatible enzymatic assay for RGS-accelerated GDP production was developed. This assay was used to screen the NIH Molecular Libraries Small Molecule Repository (MLSMR) in order to identify a small molecule that inhibits RGS4.

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: DA030555-01
Assay Submitter (PI): David Siderovski, University of North Carolina Chapel Hill (School of Medicine)
Protocol
Two microliters of 500nM (2x working concentration) is dispensed into black medium-binding solid-bottom Kalypsys plates. Then with the Kalypsys pintool 23nl of control and compounds are transferred; the control compound is 100mM CCG-4986 (working concentration). After a 10 minute incubation at room temperature 2ul of antibody is added with the Kalypsys dispenser; (-)RGS4/Transcreener GDP solution in columns 3 and 4 which consisted of 10nM (2x working concentration) Alexa633, 20uM GTP, and 20 ug/ml GDP antibody and (+)RGS4/Transcreener GDP solution in columns 1,2, and 5 to 48 which consists of 900nM RGS4 (2x working concentration), 10nM Alexa633, 20 ug/ml GDP antibody. The plate is centrifuged at 1000 RPM for 15 seconds to remove any possible bubbles and then incubated for 5 minutes at ambient room temperature. An initial fluorescent polarization read (620 nm/688 nm) is performed on the Envision before the plate is incubated for 90 minutes at 30 deg C at a relative humidity of 80%. The plate is once more centrifuged at 1000 RPM for 15 seconds and another Envision fluorescent polarization (620 nm/688 nm) read is carried out.
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 inhibitors are ranked higher than compounds that showed apparent activation.
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
5Curve_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
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_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
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.00366 uM (0.00365927μM**)% Activity at given concentration.Float%
15Activity at 0.091 uM (0.0914072μM**)% Activity at given concentration.Float%
16Activity at 0.457 uM (0.457035μM**)% Activity at given concentration.Float%
17Activity at 2.289 uM (2.28892μM**)% Activity at given concentration.Float%
18Activity at 2.860 uM (2.86μM**)% Activity at given concentration.Float%
19Activity at 5.710 uM (5.71μM**)% Activity at given concentration.Float%
20Activity at 9.139 uM (9.13936μM**)% Activity at given concentration.Float%
21Activity at 11.42 uM (11.4174μM**)% Activity at given concentration.Float%
22Activity at 19.35 uM (19.3494μM**)% Activity at given concentration.Float%
23Activity at 28.60 uM (28.6μM**)% Activity at given concentration.Float%
24Activity at 56.94 uM (56.9434μM**)% Activity at given concentration.Float%
25Activity at 80.69 uM (80.6924μM**)% Activity at given concentration.Float%
26Activity at 113.8 uM (113.83μM**)% Activity at given concentration.Float%
27Activity at 162.0 uM (161.981μM**)% Activity at given concentration.Float%
28Activity at 229.0 uM (229μM**)% Activity at given concentration.Float%
29Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: DA030555

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