Current Chemical Genomics and Translational Medicine

2009, 3 : 42-49
Published online 2009 June 16. DOI: 10.2174/1875397300903010042
Publisher ID: CCGTM-3-42

RESEARCH ARTICLE
 A High Throughput Assay to Identify Small Molecule Modulators of Prostatic Acid Phosphatase

Rylan S Larsen1 , Mark J Zylka1 and John E Scott, *,2
1 Department of Cell and Molecular Physiology, UNC Neuroscience Center, University of North Carolina at Chapel Hill, CB#7545, Chapel Hill, NC 27599, USA
2 Department of Pharmaceutical Sciences, Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, 1801 Fayetteville Street, Durham, NC 27707, USA

* Address correspondence to this author at the Department of Pharmaceutical Sciences, Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, 1801 Fayetteville Street, Durham, NC 27707, USA; E-mail: jscott@nccu.edu

ABSTRACT

Prostatic acid phosphatase (PAP) is expressed in nociceptive neurons and functions as an ectonucleotidase. Injection of the secretory isoform of PAP has potent antinociceptive effects in mouse models of chronic pain. These data suggested that a small molecule activator of PAP may have utility as a novel therapeutic for chronic pain, while inhibitors could be used to acutely inhibit PAP in vitro and in vivo. To identify small molecule modulators of PAP activity, we validated a high throughput, fluorescence-based biochemical assay and then used this assay to screen a compound library. We decreased the frequency of false positive activators by subtracting compound fluorescence from the final assay fluorescence. This approach significantly reduced the number of false positive activators found in the screen. While no activators were confirmed, seven novel inhibitors of PAP were identified. Our results suggest this high throughput assay could be used to identify small molecule modulators of PAP activity.