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 ..
BioActive Compounds: 20
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
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.
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.
* Activity Concentration. ** Test Concentration.
Data Table (Concise)