Bookmark and Share
BioAssay: AID 720504

qHTS for Inhibitors of PLK1-PDB (polo-like kinase 1 - polo-box domain): Primary Screen

The polo-like kinase 1 (Plk1) is a member of a conserved subfamily of serine / threonine protein kinases that plays a pivotal roles in cell proliferation. It has a C-terminal non-catalytic region called the polo-box domain (PBD) that plays a crucial role in the subcellular localization of the Plk1. Recent publications also revealed that Plk1 is the major kinase required for the viability of more ..
_
   
 Tested Compounds
 Tested Compounds
All(364053)
 
 
Active(10182)
 
 
Inactive(340383)
 
 
Inconclusive(13556)
 
 
 Tested Substances
 Tested Substances
All(364721)
 
 
Active(10204)
 
 
Inactive(340948)
 
 
Inconclusive(13569)
 
 
AID: 720504
Data Source: NCGC (PLK1100)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2013-07-09

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 10182
Depositor Specified Assays
AIDNameTypeComment
720510qHTS for Inhibitors of PLK1-PDB (polo-like kinase 1 - polo-box domain): Summarysummary
Description:
The polo-like kinase 1 (Plk1) is a member of a conserved subfamily of serine / threonine protein kinases that plays a pivotal roles in cell proliferation. It has a C-terminal non-catalytic region called the polo-box domain (PBD) that plays a crucial role in the subcellular localization of the Plk1. Recent publications also revealed that Plk1 is the major kinase required for the viability of activated Ras or inactivated p53 mutation-bearing cancer cells, but not the respective normal cells. Furthermore, depletion of the overexpressed Plk1 by RNAi efficiently induces apoptotic cell death in cancer and reduces tumor growth in mouse xenograft models. Thus, targeting Plk1 is an appealing strategy for anti-cancer therapy.

Therefore, this project aimed to screen the Molecular Libraries Small Molecule Repository (MLSMR) using a novel approach to find inhibitors that target the Plk1 PBD instead of the conventional N-terminal catalytic domain. This project employed a fluorescence polarization (FP) assay using a FITC-conjugated phospho-9-mer (p-9-mer) peptide and purified human Plk1 PBD protein. The identified small molecule inhibitors of Plk1 PBD can then further be developed into useful tools for biomedical and clinical research that complements or replaces current relevant disease therapy.

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

MLPCN Grant: MH100728
Assay Submitter (PI): Kyung Lee, NCI
Protocol
Four microliter of 25 nM FITC-p-9-mer + 60 nM Plk1 PBD in buffer solution (final concentrations 25 nM and 60 nM respectively) or 4 uL of 25 nM FITC-p-9-mer (final concentration 25 nM) in buffer will be dispensed into a 1,536 Greiner medium-binding black solid plate. Small molecules at final concentrations of 18 nM to 57 uM and positive control phospho-9-mer (sequence: Ac-PPLHSpTAI-NH2) at final concentrations 26 nM to 57 uM and phospho-13-mer (sequence: Ac-CETFDPPLHSpTAI-NH2) at final concentrations of 1 nM to 2.3 uM will be pin-transferred (23 nL) to the plate via a Kalypsys pin-tool robotics equipped with a 1,536-pin array. The reaction plate will be centrifuged for 1 min at 1000 rpm and incubated at room temperature for 10 min. The FITC fluorescence signals will be read using the PerkinElmer ViewLux plate reader at 480 / 20 nm excitation and 540 / 20 nm S and P polarization emission wavelengths. Reagent bottles will be kept submerged into 4 oC recirculating chiller bath and all liquid lines will be covered with aluminum foil to minimize degradation. Library plates will be screened starting from the lowest to the highest concentrations to minimize compound carryover. Vehicle only plates, with DMSO being pin-transferred instead of compound solutions, will be included regularly throughout the screen to record any systematic shifts in assay signal.
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
5Activity_ScoreActivity score.Integer
6Curve_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
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_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
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0000366000 uM (3.66e-05μM**)% Activity at given concentration.Float%
16Activity at 0.0000731000 uM (7.31e-05μM**)% Activity at given concentration.Float%
17Activity at 0.0001870464 uM (0.000187046μM**)% Activity at given concentration.Float%
18Activity at 0.0003360693 uM (0.000336069μM**)% Activity at given concentration.Float%
19Activity at 0.0007304503 uM (0.00073045μM**)% Activity at given concentration.Float%
20Activity at 0.00149 uM (0.00148767μM**)% Activity at given concentration.Float%
21Activity at 0.00218 uM (0.00217569μM**)% Activity at given concentration.Float%
22Activity at 0.00370 uM (0.00369918μM**)% Activity at given concentration.Float%
23Activity at 0.00833 uM (0.00833304μM**)% Activity at given concentration.Float%
24Activity at 0.018 uM (0.017974μM**)% Activity at given concentration.Float%
25Activity at 0.032 uM (0.0323431μM**)% Activity at given concentration.Float%
26Activity at 0.049 uM (0.0490185μM**)% Activity at given concentration.Float%
27Activity at 0.093 uM (0.0927383μM**)% Activity at given concentration.Float%
28Activity at 0.206 uM (0.206057μM**)% Activity at given concentration.Float%
29Activity at 0.449 uM (0.449185μM**)% Activity at given concentration.Float%
30Activity at 0.785 uM (0.785224μM**)% Activity at given concentration.Float%
31Activity at 1.205 uM (1.20533μM**)% Activity at given concentration.Float%
32Activity at 2.302 uM (2.30206μM**)% Activity at given concentration.Float%
33Activity at 5.061 uM (5.06113μM**)% Activity at given concentration.Float%
34Activity at 11.18 uM (11.1773μM**)% Activity at given concentration.Float%
35Activity at 19.62 uM (19.6209μM**)% Activity at given concentration.Float%
36Activity at 26.11 uM (26.1079μM**)% Activity at given concentration.Float%
37Activity at 57.27 uM (57.2671μM**)% Activity at given concentration.Float%
38Activity at 114.6 uM (114.624μM**)% Activity at given concentration.Float%
39Activity at 198.8 uM (198.755μM**)% Activity at given concentration.Float%
40Activity at 304.0 uM (304μM**)% Activity at given concentration.Float%
41Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH100728

Data Table (Concise)
Classification
PageFrom: