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BioAssay: AID 2710

Kinase Inhibition Profile Study on Inhibitors of CDC-like Kinase 4

Alternative splicing of pre-mRNAs plays a major role in the regulation of eukaryotic gene expression. Alternative splicing is now thought to be one of the primary reasons for the complexity of higher organisms leading to an average of three protein isoforms per gene. Aberrant splicing can lead to a number of diseases and is known to play a role in rare genetic diseases such as progeria syndrome. more ..
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 Tested Compounds
 Tested Compounds
All(28)
 
 
Active(28)
 
 
 Tested Substances
 Tested Substances
All(28)
 
 
Active(28)
 
 
AID: 2710
Data Source: NCGC (CLK112)
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-03-26
Modify Date: 2010-10-21

Data Table ( Complete ):           Active    All
Targets
Sequence: CDC-like kinase 4 [Homo sapiens]
Description ..   
Protein Family: Protein Kinases, catalytic domain

Gene:CLK4     Related Protein 3D Structures     More BioActivity Data..


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BioActive Compounds: 28
Depositor Specified Assays
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AIDNameTypeProbeComment
1459Validation of Assay for Modulators of Lamin A Splicingconfirmatory
1487qHTS Assay for Modulators of Lamin A Splicingconfirmatory
1498Confirmation Concentration-Response Assay for Modulators of Lamin A Splicingconfirmatory
1770qHTS Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)confirmatory
1970Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay)confirmatory
1997qHTS for Inhibitors of CDC-like Kinase 4: Summarysummary3
488872qHTS for Inhibitors of DYRK1A: Summarysummary1
488883qHTS for Inhibitors of DYRK1B: Summarysummary1
488887qHTS for Inhibitors of CDC-like Kinase 1 and 4: Summarysummary1
493204Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay): SAR round 2confirmatory
Description:
NIH Molecular Libraries Probe Production Centers Network [MLPCN]
NIH Chemical Genomics Center [NCGC]
MLSCN Grant: 1R03MH084827-01
Assay Submitter (PI): Tom Misteli

NCGC Assay Overview:

Alternative splicing of pre-mRNAs plays a major role in the regulation of eukaryotic gene expression. Alternative splicing is now thought to be one of the primary reasons for the complexity of higher organisms leading to an average of three protein isoforms per gene. Aberrant splicing can lead to a number of diseases and is known to play a role in rare genetic diseases such as progeria syndrome. Hutchinson-Gilford Progeria Syndrome (HGPS) is a pediatric premature aging disease caused by a spontaneous mutation in the lamin A/C (LMNA) gene due to a single point mutation in the lamin A gene effecting 1 in 4 million people.

To identify both specific splicing modulators of the lamin A gene as well as general splicing modulators, a cell-based HTS assay was developed using a minigene reporter system (AIDs, 1459 & 1487). A quinazoline (CID: 16759567) showed weak activity as an inhibitor of aberrant splicing using the minigene reporter system (AID 1498). A preliminary kinase profile of this compound showed activity in splicing related kinases belonging to the CDC-like kinase (Clk) family particularly the Clk4. The Clk family is part of the CMGC group of protein kinases which includes Dyrks and 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 [2] and Alzheimer disease [3]. This AID summarizes the characterization of lead Clk4 inhibitors in a panel of these related kinases: Clk1, 2 & 3, DYRK1/DYRK1A, and DYRK1B. Potency against Clk4 was also confirmed in the panel.
Protocol
Assay Protocol:
Compound IC50s were determined at Reaction Biology (http://www.reactionbiology.com) with ten concentration points starting at 10 uM in the assay using a 3-fold dilution series. The control compound staurosporine was also tested in the 10-dose IC50 mode with the same 3-fold serial dilution, again starting at 10 uM. All kinase reactions were performed using 10 uM ATP in the assay. 100% activity was taken from kinase reactions with DMSO alone (the vehicle used for compound stock solutions).
Comment
Compounds with the potency below 1uM for Clk4 are given a score of 80, the rest of them are give a score of 50.
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed based on the compound activity across various assays tested.String
2Potency_Clk4The concentration of sample yielding half-maximal inhibition in Clk4 assay.FloatμM
3Potency_Clk1The concentration of sample yielding half-maximal inhibition in Clk1 assay.FloatμM
4Potency_Clk2The concentration of sample yielding half-maximal inhibition in Clk2 assay.FloatμM
5Potency_Clk3The concentration of sample yielding half-maximal inhibition in Clk3 assay.FloatμM
6Potency_Dyrk1aThe concentration of sample yielding half-maximal inhibition in Dyrk1a assay.FloatμM
7Potency_Dyrk1bThe concentration of sample yielding half-maximal inhibition in Dyrk1b assay.FloatμM
Additional Information
Grant Number: 1R03MH084827-01

Data Table (Concise)
Classification
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