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BioAssay: AID 1477

qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia

Core Binding Factor (CBF) abnormalities are associated with 20-25% of all acute myeloid leukemias (AML), of which 5-10% are further sub classified as acute myelomonocytic leukemia with eosinophilia, also known as M4Eo in the FAB classification scheme. This subtype of leukemia is usually associated with chromosome 16 inversion (p13:q22). Chromosome 16 inversion results in formation of the more ..
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 Tested Compounds
 Tested Compounds
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Active(3)
 
 
Inactive(223597)
 
 
Inconclusive(7190)
 
 
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 Tested Substances
All(234798)
 
 
Active(3)
 
 
Inactive(227513)
 
 
Inconclusive(7282)
 
 
AID: 1477
Data Source: NCGC (CBFB001)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2009-01-02
Modify Date: 2011-03-21

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 3
Related Experiments
AIDNameTypeComment
1484Quantitative High-Throughput Screen for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: SummarySummarydepositor-specified cross reference
504370qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: ME-1 cellular proliferationConfirmatorydepositor-specified cross reference
504371qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: ME-1 cellular proliferation, SAR for ProbeConfirmatorydepositor-specified cross reference
504373qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: Zebrafish assay for Hematopoiesis, SAR for ProbeOtherdepositor-specified cross reference
504374qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: Hit ValidationConfirmatorydepositor-specified cross reference
504375qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: Hit Validation with Orthogonal AssayConfirmatorydepositor-specified cross reference
504378qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: SAR for Probe with Orthogonal AssayConfirmatorydepositor-specified cross reference
504386qHTS Assay for Compounds Blocking the Interaction Between CBF-beta and RUNX1 for the Treatment of Acute Myeloid Leukemia: SAR for ProbeConfirmatorydepositor-specified cross reference
Description:
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Production centers Network [MLPCN]

MLPCN Grant: X01MH083259-01
Assay Submitter (PI): Paul Liu

NCGC Assay Overview:

Core Binding Factor (CBF) abnormalities are associated with 20-25% of all acute myeloid leukemias (AML), of which 5-10% are further sub classified as acute myelomonocytic leukemia with eosinophilia, also known as M4Eo in the FAB classification scheme. This subtype of leukemia is usually associated with chromosome 16 inversion (p13:q22). Chromosome 16 inversion results in formation of the fusion oncogene CBFB-MYH11, which encodes the fusion protein CBF-beta-SMMHC. This fusion protein binds to RUNX1 (AML1) with high affinity and dominantly inhibits RUNX1 function preventing definitive hematopoiesis.

It has been hypothesized that the interference of CBF-beta and RUNX1 binding could have therapeutic implications for patients with CBF mediated leukemias. We developed a CBF-beta and RUNX1 binding assay in AlphaScreenTM format and optimized it for high throughput screening to search for inhibitors. The goal of this project is to screen MLPCN's compound collection (MLSMR) for identifying the inhibitors of the interaction between CBF-beta and RUNX1 proteins as research probes. If these probes work well in the animal models, the ultimate goal of this project is to development a new drug treatment for this disease.

This assay can not be used to assess how apparent inhibitors interfere with protein binding, and whether inhibitors specifically bind to either protein. Disruption of this protein-protein interaction, through any mechanism, is the intended target activity.
Protocol
NCGC Assay Protocol Summary:

Step, Parameter, Description
(1) Reagent, 3 uL CBFb-His/RUNX1-biotin mix incubated together for 20 minutes at room temperature (30 nM each)
(2) Compound or control, 23 nL compound stock solution
(3) Incubation, 15 minutes at Room temperature
(4) Reagent, 1 uL 50 ng each of beads (Nickel beads and streptavidin beads)
(5) Incubation, 90 minutes at Room temperature
(6) Detection, Excitation=365 nm Emission=440 nm, Envision reader in AlphaScreenTM mode

Keywords: MLSMR, MLPCN, NIH Roadmap, qHTS, NCGC, Core binding factor, CBF, CBF-beta, Acute myeloid leukemia
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: Binding
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 signficant, 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 0.00164 uM (0.00164368μM**)% Activity at given concentration.Float%
15Activity at 0.00368 uM (0.00367816μM**)% Activity at given concentration.Float%
16Activity at 0.00822 uM (0.00821839μM**)% Activity at given concentration.Float%
17Activity at 0.018 uM (0.0183908μM**)% Activity at given concentration.Float%
18Activity at 0.041 uM (0.0411494μM**)% Activity at given concentration.Float%
19Activity at 0.093 uM (0.093252μM**)% Activity at given concentration.Float%
20Activity at 0.133 uM (0.133162μM**)% Activity at given concentration.Float%
21Activity at 0.202 uM (0.201713μM**)% Activity at given concentration.Float%
22Activity at 0.417 uM (0.417308μM**)% Activity at given concentration.Float%
23Activity at 0.514 uM (0.513579μM**)% Activity at given concentration.Float%
24Activity at 0.837 uM (0.836802μM**)% Activity at given concentration.Float%
25Activity at 1.286 uM (1.28634μM**)% Activity at given concentration.Float%
26Activity at 2.359 uM (2.35902μM**)% Activity at given concentration.Float%
27Activity at 3.436 uM (3.43578μM**)% Activity at given concentration.Float%
28Activity at 5.174 uM (5.17391μM**)% Activity at given concentration.Float%
29Activity at 10.68 uM (10.6773μM**)% Activity at given concentration.Float%
30Activity at 13.09 uM (13.0858μM**)% Activity at given concentration.Float%
31Activity at 21.72 uM (21.7182μM**)% Activity at given concentration.Float%
32Activity at 32.14 uM (32.1375μM**)% Activity at given concentration.Float%
33Activity at 59.83 uM (59.8347μM**)% Activity at given concentration.Float%
34Activity at 89.08 uM (89.0793μM**)% Activity at given concentration.Float%
35Activity at 135.5 uM (135.538μM**)% Activity at given concentration.Float%
36Activity at 187.4 uM (187.356μM**)% 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: X01MH083259-01

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