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BioAssay: AID 504937

Inhibitors of Secretory Acid Sphingomyelinase (S-ASM): qHTS

Atherosclerosis is the underlying process for cardiovascular disease and thus is a major contributor to disease burden in the population. The infiltration of LDL into the subendothelial space of the vessel wall, and more importantly its retention, has been shown to be the key factors in the initiation of atherosclerosis. Recently, it has been found that secretory acid sphingomyelinase (S-ASM) more ..
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 Tested Compounds
 Tested Compounds
All(337893)
 
 
Active(20)
 
 
Inactive(335359)
 
 
Inconclusive(2515)
 
 
 Tested Substances
 Tested Substances
All(338135)
 
 
Active(20)
 
 
Inactive(335597)
 
 
Inconclusive(2518)
 
 
AID: 504937
Data Source: NCGC (ASM101)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2011-07-08
Modify Date: 2012-05-30

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 20
Related Experiments
AIDNameTypeComment
504940Inhibitors of Secretory Acid Sphingomyelinase (S-ASM): SummarySummarydepositor-specified cross reference
624178qHTS for Inhibitors of Human Acid Sphingomyelinase Assay: Native SubstrateConfirmatorydepositor-specified cross reference
Description:
Atherosclerosis is the underlying process for cardiovascular disease and thus is a major contributor to disease burden in the population. The infiltration of LDL into the subendothelial space of the vessel wall, and more importantly its retention, has been shown to be the key factors in the initiation of atherosclerosis. Recently, it has been found that secretory acid sphingomyelinase (S-ASM) action on LDL promotes the retention of LDL by causing its aggregation and enhancing its uptake by macrophages. Thus, selective small molecule inhibitors of S-ASM should block LDL modification and reduce aortic LDL retention and may serve as new agents for the treatment of atherosclerosis.

In a collaboration between National Heart, Lung, and Blood Institute and the NCGC a high-throughput amenable screen was developed to identify potent and selective small molecule inhibitors of S-ASM. This enzymatic, fluorescent screen uses an ASM enzyme preparation derived from human placenta and was tested against the NIH Molecular Libraries Small Molecule Repository (MLSMR).

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

MLPCN Grant: MH093173-01
Assay Submitter (PI): Alan Remaley, National Heart, Lung, and Blood Institute and Wei Zhen, NIH Chemical Genomics Center
Protocol
Two microliters of ASM enzyme (1:150 dilution from stock) are dispensed in 1536-well plate. 23 nL of compound and control are pinned into assay plate using the Kalypsys pintool. The plates are then incubated for 10 minutes at ambient room temperature. Then, 2 uL of the flurogenic substrate, HMU-PC0, (9 uM final) are added to the plate. An incubation for 60 minutes takes place at 37 masculineC. After that, 4 uL of stop solution is added to raise the pH to 10 to end the kinetic reaction. Plate is read on ViewLux, with detection set at Ex = 386 (10) nm and Em = 447 (10) nm.
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
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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 0.460 uM (0.46μM**)% Activity at given concentration.Float%
15Activity at 2.300 uM (2.3μM**)% Activity at given concentration.Float%
16Activity at 11.50 uM (11.5μM**)% Activity at given concentration.Float%
17Activity at 57.50 uM (57.5μM**)% Activity at given concentration.Float%
18Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH093173

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