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BioAssay: AID 588579

qHTS for Inhibitors of Polymerase Kappa

The processing and repair of DNA interstrand crosslinks (ICL) in eukaryotic cells are extremely complex, potentially involving multiple DNA repair and damage tolerance pathways, including homologous recombination, nucleotide excision repair, translesion synthesis (TLS), and cell cycle checkpoints. In some repair models, the dually-incised strand possesses sufficient mobility that a DNA polymerase more ..
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 Tested Compounds
 Tested Compounds
All(395291)
 
 
Active(2035)
 
 
Inactive(388558)
 
 
Inconclusive(5142)
 
 
 Tested Substances
 Tested Substances
All(404211)
 
 
Active(2159)
 
 
Inactive(396841)
 
 
Inconclusive(5211)
 
 
AID: 588579
Data Source: NCGC (PolK100)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-10-20

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 2035
Related Experiments
AIDNameTypeComment
588638qHTS for Inhibitors of Polymerase Kappa: SummarySummarydepositor-specified cross reference
720501qHTS for Inhibitors of Polymerase Kappa: Confirmatory Assay for Cherry-picked CompoundsConfirmatorydepositor-specified cross reference
Description:
The processing and repair of DNA interstrand crosslinks (ICL) in eukaryotic cells are extremely complex, potentially involving multiple DNA repair and damage tolerance pathways, including homologous recombination, nucleotide excision repair, translesion synthesis (TLS), and cell cycle checkpoints. In some repair models, the dually-incised strand possesses sufficient mobility that a DNA polymerase can strand displace the nucleotide patch that is 5' to the lesion, then replicate past the ICL site to complete the repair gap-filling synthesis. Polymerase kappa has been implicated in this project and is hence of interest.

To that end, in a collaboration between the Oregon Health & Science University and the NIH Chemical Genomics Center a 1536-well high-throughput screen was developed. This fluorescent screen was assayed against the NIH Molecular Libraries Small Molecule Repository (MLSMR) to discover inhibitors of Pol Kappa.


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

MLPCN Grant: MH094179
Assay Submitter (PI): Stephen Lloyd, Oregon Health & Science University
Protocol
Three microliters of reagents (buffer in column 3 and 4 as negative control and 10 nM Pol kappa in columns 1, 2, and 5-48) were dispensed into a 1536-well black solid-bottom plate. Compounds (23 nL) were transferred via Kalypsys pin tool equipped with 1536-pin array. The plates were then incubated for 15 min at room temperature, and 1 uL substrate (50 nM final concentration) were then added to start the reaction and kinetically read twice at 0 min and 10 min on the Viewlux 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 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
Show more
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 0.0003270000 uM (0.000327μM**)% Activity at given concentration.Float%
15Activity at 0.0007732774 uM (0.000773277μM**)% Activity at given concentration.Float%
16Activity at 0.00163 uM (0.00163499μM**)% Activity at given concentration.Float%
17Activity at 0.00369 uM (0.00369141μM**)% Activity at given concentration.Float%
18Activity at 0.00818 uM (0.00817833μM**)% Activity at given concentration.Float%
19Activity at 0.020 uM (0.0200781μM**)% Activity at given concentration.Float%
20Activity at 0.030 uM (0.0298882μM**)% Activity at given concentration.Float%
21Activity at 0.047 uM (0.0470036μM**)% Activity at given concentration.Float%
22Activity at 0.101 uM (0.101257μM**)% Activity at given concentration.Float%
23Activity at 0.151 uM (0.150818μM**)% Activity at given concentration.Float%
24Activity at 0.243 uM (0.242831μM**)% Activity at given concentration.Float%
25Activity at 0.477 uM (0.476631μM**)% Activity at given concentration.Float%
26Activity at 0.759 uM (0.758663μM**)% Activity at given concentration.Float%
27Activity at 1.287 uM (1.28744μM**)% Activity at given concentration.Float%
28Activity at 2.393 uM (2.39272μM**)% Activity at given concentration.Float%
29Activity at 3.818 uM (3.81766μM**)% Activity at given concentration.Float%
30Activity at 6.336 uM (6.33552μM**)% Activity at given concentration.Float%
31Activity at 11.99 uM (11.9931μM**)% Activity at given concentration.Float%
32Activity at 19.37 uM (19.3713μM**)% Activity at given concentration.Float%
33Activity at 31.37 uM (31.366μM**)% Activity at given concentration.Float%
34Activity at 60.11 uM (60.1052μM**)% Activity at given concentration.Float%
35Activity at 107.2 uM (107.163μM**)% Activity at given concentration.Float%
36Activity at 158.4 uM (158.401μM**)% Activity at given concentration.Float%
37Activity at 229.0 uM (229μM**)% Activity at given concentration.Float%
38Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH094179

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