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BioAssay: AID 588590

qHTS for Inhibitors of Polymerase Iota

DNA polymerases, such as Pol iota, are specifically involved in DNA repair. Pol iota specifically plays an important role in translesion synthesis, where the normal high-fidelity DNA polymerases cannot proceed and DNA synthesis stalls. It has been reported that human pol iota may be upregulated in a number of human cancers and specifically during the progression of breast cancer, indicating that more ..
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 Tested Compounds
 Tested Compounds
All(386057)
 
 
Active(3988)
 
 
Inactive(354454)
 
 
Inconclusive(28338)
 
 
 Tested Substances
 Tested Substances
All(391277)
 
 
Active(4029)
 
 
Inactive(358790)
 
 
Inconclusive(28458)
 
 
AID: 588590
Data Source: NCGC (PolI100)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-10-20

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 3988
Depositor Specified Assays
AIDNameTypeComment
588623qHTS for Inhibitors of Polymerase Iota: Summarysummary
720496qHTS for Inhibitors of Polymerase Iota: Confirmatory Assay for Cherry-picked Compoundsconfirmatory
Description:
DNA polymerases, such as Pol iota, are specifically involved in DNA repair. Pol iota specifically plays an important role in translesion synthesis, where the normal high-fidelity DNA polymerases cannot proceed and DNA synthesis stalls. It has been reported that human pol iota may be upregulated in a number of human cancers and specifically during the progression of breast cancer, indicating that pol iota's activities within normal living cells must be tightly regulated. Thus, while pol eta's (and most likely iota's) normal function is to protect humans against the deleterious consequences of DNA damage, under certain conditions they can have deleterious effects on human health. As a consequence, we propose to utilize a high throughput replication assay to identify small molecule inhibitors of pol iota.

In a collaboration between the National Institute of Child Health & Human Development (NICHD) and NIH Chemical Genomics Center, a high-throughput, fluorescent screen was developed to screen the NIH Molecular Libraries Small Molecule Repository (MLSMR). This screen is used to identify inhibitors of pol iota.

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

MLPCN Grant: MH090825
Assay Submitter (PI): Roger Woodgate, NICHD)
Protocol
Three microliters of reagents (buffer in column 3 and 4 as negative control and 10 nM Pol iota in columns 1, 2, and 5-48) will be dispensed into 1,536-well black solid-bottomed plate. Compounds (23 nL) will be transferred via Kalypsys pin tool equipped with 1536-pin array. The plates will then be incubated for 15 min at room temperature, and 1 muL substrate (50 nM final concentration) will be 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.
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 0.00174 uM (0.00174286μM**)% Activity at given concentration.Float%
15Activity at 0.00357 uM (0.00357179μM**)% Activity at given concentration.Float%
16Activity at 0.00697 uM (0.00697143μM**)% Activity at given concentration.Float%
17Activity at 0.016 uM (0.0159735μM**)% Activity at given concentration.Float%
18Activity at 0.028 uM (0.0278857μM**)% Activity at given concentration.Float%
19Activity at 0.056 uM (0.0557714μM**)% Activity at given concentration.Float%
20Activity at 0.105 uM (0.105103μM**)% Activity at given concentration.Float%
21Activity at 0.226 uM (0.225908μM**)% Activity at given concentration.Float%
22Activity at 0.447 uM (0.447463μM**)% Activity at given concentration.Float%
23Activity at 0.627 uM (0.627293μM**)% Activity at given concentration.Float%
24Activity at 0.951 uM (0.95053μM**)% Activity at given concentration.Float%
25Activity at 1.818 uM (1.8177μM**)% Activity at given concentration.Float%
26Activity at 2.333 uM (2.33278μM**)% Activity at given concentration.Float%
27Activity at 4.073 uM (4.0734μM**)% Activity at given concentration.Float%
28Activity at 6.884 uM (6.8844μM**)% Activity at given concentration.Float%
29Activity at 11.29 uM (11.2905μM**)% Activity at given concentration.Float%
30Activity at 15.41 uM (15.4073μM**)% Activity at given concentration.Float%
31Activity at 25.59 uM (25.5875μM**)% Activity at given concentration.Float%
32Activity at 50.19 uM (50.1855μM**)% Activity at given concentration.Float%
33Activity at 58.90 uM (58.9041μM**)% Activity at given concentration.Float%
34Activity at 114.8 uM (114.764μM**)% Activity at given concentration.Float%
35Activity at 162.0 uM (161.981μM**)% Activity at given concentration.Float%
36Activity at 229.0 uM (229μM**)% Activity at given concentration.Float%
37Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH090825

Data Table (Concise)
Classification
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