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BioAssay: AID 2705

Assay for Inhibitors of Dual-Specificity Tyrosine-(Y)-Phosphorylation Regulated Kinase 1A (Kinase-Glo assay)

The CMGC group of protein kinases which includes CDK-like kinases (Clk) and Dyks are essential kinases found in all eukaryotes. Clks have been implicated to play a role in the regulation of alternative splicing [1]; while Dyrks, in particular over expression of Dyrk1A, has been linked to diseases as Down syndrome (DS; OMIM 190685) [2] and Alzheimer disease (AD; OMIM 104300) [3]. Among the more ..
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 Tested Compounds
 Tested Compounds
All(101)
 
 
Active(87)
 
 
Inactive(4)
 
 
Inconclusive(10)
 
 
 Tested Substances
 Tested Substances
All(101)
 
 
Active(87)
 
 
Inactive(4)
 
 
Inconclusive(10)
 
 
AID: 2705
Data Source: NCGC (CLK202)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-03-26
Modify Date: 2010-10-21

Data Table ( Complete ):           View Active Data    View All Data
Target
Sequence: dual specificity tyrosine-phosphorylation-regulated kinase 1A isoform 1 [Homo sapiens]
Description ..   
Protein Family: Protein Kinases, catalytic domain

Gene:DYRK1A     Related Protein 3D Structures     More BioActivity Data..
BioActive Compounds: 87
Related Experiments
Show more
AIDNameTypeProbeComment
1770qHTS Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)Confirmatory depositor-specified cross reference
1997qHTS for Inhibitors of CDC-like Kinase 4: SummarySummary3 depositor-specified cross reference
488872qHTS for Inhibitors of DYRK1A: SummarySummary1 depositor-specified cross reference
488883qHTS for Inhibitors of DYRK1B: SummarySummary1 depositor-specified cross reference
488887qHTS for Inhibitors of CDC-like Kinase 1 and 4: SummarySummary1 depositor-specified cross reference
493206Assay for Inhibitors of Dual-Specificity Tyrosine-(Y)-Phosphorylation Regulated Kinase 1A (Kinase-Glo assay): round 2 SARConfirmatory depositor-specified cross reference
1771qHTS Assay for Inhibitors of CDC-like Kinase 4 (ADP-Glo Assay)Confirmatory same project related to Summary assay
1795qHTS Assay for Inhibitors of CDC-like Kinase 4 (ADP-FP Assay)Confirmatory same project related to Summary assay
1969Confirmaiton Assay for Inhibitors of CDC-like Kinase 4 (ADP-Glo Assay)Confirmatory same project related to Summary assay
1970Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)Confirmatory same project related to Summary assay
1983Confirmation Assay for Inhibitors of CDC-like Kinase 4 (ADP-FP Assay)Confirmatory same project related to Summary assay
2710Kinase Inhibition Profile Study on Inhibitors of CDC-like Kinase 4Other same project related to Summary assay
493204Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay): SAR round 2Confirmatory same project related to Summary assay
504421Kinase Inhibition Study on Inhibitors of CDC-like Kinase 4 (Reaction Biology data): SAR round 2Confirmatory same project related to Summary assay
504424Kinase Inhibition Study on Inhibitors of Dyrk1a (Reaction Biology data)Confirmatory same project related to Summary assay
504427Kinase Inhibition Study on Inhibitors of CDC-like Kinase 2 (Reaction Biology data)Confirmatory same project related to Summary assay
504428Kinase Inhibition Study on Inhibitors of CDC-like Kinase 3 (Reaction Biology data)Confirmatory same project related to Summary assay
504429Kinase Inhibition Study on Inhibitors of Dyrk1b (Reaction Biology data)Confirmatory same project related to Summary assay
504430Kinase Inhibition Study on Inhibitors of CDC-like Kinase 1 (Reaction Biology data)Confirmatory same project related to Summary assay
504534Secondary Assay for Inhibitors of Human Cdc-like kinase 4 (Clk4): HEK-293 Cell Viability AssayConfirmatory same project related to Summary assay
624034Reaction Biology data for inhibitors of CLK4: Probe SARConfirmatory same project related to Summary assay
624045Reaction Biology data for inhibitors of DYRK1A: Probe SARConfirmatory same project related to Summary assay
624046Reaction Biology data for inhibitors of DYRK1B: Probe SARConfirmatory same project related to Summary assay
624047Reaction Biology data for inhibitors of CLK1: Probe SARConfirmatory same project related to Summary assay
624048Reaction Biology data for inhibitors of CLK2: Probe SARConfirmatory same project related to Summary assay
624049Reaction Biology data for inhibitors of CLK3: Probe SARConfirmatory same project related to Summary assay
624093Confirmation Assay for Inhibitors of Dyrk1a (Kinase-Glo Assay): KU probeConfirmatory same project related to Summary assay
624094Confirmation Assay for Inhibitors of Clk4 (Kinase-Glo Assay): KU probeConfirmatory same project related to Summary assay
1459Validation of Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1487qHTS Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1498Confirmation Concentration-Response Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1970Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)Confirmatory same project related to Summary assay
2710Kinase Inhibition Profile Study on Inhibitors of CDC-like Kinase 4Other same project related to Summary assay
493204Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay): SAR round 2Confirmatory same project related to Summary assay
504421Kinase Inhibition Study on Inhibitors of CDC-like Kinase 4 (Reaction Biology data): SAR round 2Confirmatory same project related to Summary assay
504424Kinase Inhibition Study on Inhibitors of Dyrk1a (Reaction Biology data)Confirmatory same project related to Summary assay
504427Kinase Inhibition Study on Inhibitors of CDC-like Kinase 2 (Reaction Biology data)Confirmatory same project related to Summary assay
504428Kinase Inhibition Study on Inhibitors of CDC-like Kinase 3 (Reaction Biology data)Confirmatory same project related to Summary assay
504429Kinase Inhibition Study on Inhibitors of Dyrk1b (Reaction Biology data)Confirmatory same project related to Summary assay
504430Kinase Inhibition Study on Inhibitors of CDC-like Kinase 1 (Reaction Biology data)Confirmatory same project related to Summary assay
1459Validation of Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1487qHTS Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1498Confirmation Concentration-Response Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1970Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)Confirmatory same project related to Summary assay
2710Kinase Inhibition Profile Study on Inhibitors of CDC-like Kinase 4Other same project related to Summary assay
493204Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay): SAR round 2Confirmatory same project related to Summary assay
504421Kinase Inhibition Study on Inhibitors of CDC-like Kinase 4 (Reaction Biology data): SAR round 2Confirmatory same project related to Summary assay
504424Kinase Inhibition Study on Inhibitors of Dyrk1a (Reaction Biology data)Confirmatory same project related to Summary assay
504427Kinase Inhibition Study on Inhibitors of CDC-like Kinase 2 (Reaction Biology data)Confirmatory same project related to Summary assay
504428Kinase Inhibition Study on Inhibitors of CDC-like Kinase 3 (Reaction Biology data)Confirmatory same project related to Summary assay
504429Kinase Inhibition Study on Inhibitors of Dyrk1b (Reaction Biology data)Confirmatory same project related to Summary assay
504430Kinase Inhibition Study on Inhibitors of CDC-like Kinase 1 (Reaction Biology data)Confirmatory same project related to Summary assay
1459Validation of Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1487qHTS Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1498Confirmation Concentration-Response Assay for Modulators of Lamin A SplicingConfirmatory same project related to Summary assay
1970Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)Confirmatory same project related to Summary assay
2710Kinase Inhibition Profile Study on Inhibitors of CDC-like Kinase 4Other same project related to Summary assay
493204Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay): SAR round 2Confirmatory same project related to Summary assay
504421Kinase Inhibition Study on Inhibitors of CDC-like Kinase 4 (Reaction Biology data): SAR round 2Confirmatory same project related to Summary assay
504424Kinase Inhibition Study on Inhibitors of Dyrk1a (Reaction Biology data)Confirmatory same project related to Summary assay
504427Kinase Inhibition Study on Inhibitors of CDC-like Kinase 2 (Reaction Biology data)Confirmatory same project related to Summary assay
504428Kinase Inhibition Study on Inhibitors of CDC-like Kinase 3 (Reaction Biology data)Confirmatory same project related to Summary assay
504429Kinase Inhibition Study on Inhibitors of Dyrk1b (Reaction Biology data)Confirmatory same project related to Summary assay
504430Kinase Inhibition Study on Inhibitors of CDC-like Kinase 1 (Reaction Biology data)Confirmatory same project related to Summary assay
Description:
NIH Molecular Libraries Probe Production Centers Network [MLPCN]
NIH Chemical Genomics Center [NCGC]
MLSCN Grant: 1R03MH084827-01
PI Name: Tom Misteli

NCGC Assay Overview:

The CMGC group of protein kinases which includes CDK-like kinases (Clk) and Dyks are essential kinases found in all eukaryotes. Clks have been implicated to play a role in the regulation of alternative splicing [1]; while Dyrks, in particular over expression of Dyrk1A, has been linked to diseases as Down syndrome (DS; OMIM 190685) [2] and Alzheimer disease (AD; OMIM 104300) [3]. Among the members of the CMGC group, the Clks and the Dyrks are very closely related [4] therefore to determine validity and selectivity of an active compound observed in the CLK4 qHTS screen (AID:1770), a bioluminescent counter-screen against Dyrk1A was developed. In addition, potent and selective compounds against Dyrk1A were identified to further assess the role of this enzyme in DS and AD.
Dyrk1A (Invitrogen, cat # PV3785) was assayed using ATP and DYRKtide (AnaSpec, cat# 62698) as substrates. Promega Kinase-Glo (cat# V6712) technology was used to detect the residual ATP following kinetic reaction. The Kinase-Glo Plus contains Ultra-Glo [5] luciferase and D-luciferin which generates a bioluminescence signal from the remaining ATP. No enzyme and harmine (Sigma, cat # 286044-1G), a compound shown to inhibit Dyrks [6] were used as a positive control; while DMSO treatment was used as a negative control.
Protocol
NCGC Assay Protocol Summary:

Two uL/well of substrate solution (8uM Dyrktide, 2uM ATP, 25mM Tris-HCl pH 7.5, 10mM MgCl2 , 0.5mM EGTA, 5mM cystein, 0.01% Triton X-100 final concentration) was dispensed into 1536-well assay plates (Greiner, solid white, medium-binding plates) with Aurora Discovery BioRAPTR Flying Reagent Dispenser (FRD, Beckton-Dickenson). Compound solution (23 nL) was transferred to the assay plate using Kalypsys pin tool equipped with a 1536-pin tool. One uL/well enzyme solution (5nM Dyrk1A, 25mM Tris-HCl pH7.5, 10mM MgCl2, 0.5mM EGTA, 5mM cystein, 0.01% Triton X-100 final concentration) was then added using FRD yielding a total kinase reaction volume of 3uL/well. After 2 hours of room temperature incubation, 3uL Kinase-Glo detection reagent was added for a final assay volume of 6uL/well. The plates were spun for 30 seconds (1000 rpm) and incubated at ambient temperature for 2 minutes followed by a luminescence read using Perkin Elmer ViewLux plate reader at 3 seconds exposure time and 2x binning.
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.
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 0.0004342330 uM (0.000434233μM**)% Activity at given concentration.Float%
15Activity at 0.00130 uM (0.0013027μM**)% Activity at given concentration.Float%
16Activity at 0.00391 uM (0.0039081μM**)% Activity at given concentration.Float%
17Activity at 0.012 uM (0.0117243μM**)% Activity at given concentration.Float%
18Activity at 0.035 uM (0.0351729μM**)% Activity at given concentration.Float%
19Activity at 0.106 uM (0.105519μM**)% Activity at given concentration.Float%
20Activity at 0.317 uM (0.316556μM**)% Activity at given concentration.Float%
21Activity at 0.950 uM (0.949668μM**)% Activity at given concentration.Float%
22Activity at 2.849 uM (2.849μM**)% Activity at given concentration.Float%
23Activity at 8.547 uM (8.54701μM**)% Activity at given concentration.Float%
24Activity at 25.64 uM (25.641μM**)% Activity at given concentration.Float%
25Activity at 76.92 uM (76.9231μM**)% Activity at given concentration.Float%
26Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: 1R03MH084827-01

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