title: Modulators of intestinal alkaline phosphatase.
abstract: Small molecule modulators of phosphatases can lead to clinically useful drugs and serve as invaluable tools to study functional roles of various phosphatases in vivo. Here, we describe lead discovery strategies for identification of inhibitors and activators of intestinal alkaline phosphatases. To identify isozyme-selective inhibitors and activators of the human and mouse intestinal alkaline phosphatases, ultrahigh throughput chemiluminescent assays, utilizing CDP-Star as a substrate, were developed for murine intestinal alkaline phosphatase (mIAP), human intestinal alkaline phosphatase (hIAP), human placental alkaline phosphatase (PLAP), and human tissue-nonspecific alkaline phosphatase (TNAP) isozymes. Using these 1,536-well assays, concurrent HTS screens of the MLSMR library of 323,000 compounds were conducted for human and mouse IAP isozymes monitoring both inhibition and activation. This parallel screening approach led to identification of a novel inhibitory scaffold selective for murine intestinal alkaline phosphatase. SAR efforts based on parallel testing of analogs against different AP isozymes generated a potent inhibitor of the murine IAP with IC50 of 540 nM, at least 65-fold selectivity against human TNAP, and >185 selectivity against human PLAP.
authors.affiliation: Conrad Prebys Center for Chemical Genomics, Sanford-Burnham Medical Research Institute, La Jolla, CA, USA.
authors.forename: Ekaterina V
authors.initials: EV
authors.lastname: Bobkova
authors.forename: Tina
authors.initials: T
authors.lastname: Kiffer-Moreira
authors.forename: Eduard A
authors.initials: EA
authors.lastname: Sergienko
mesh_terms: D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000818:Animals; D055808:Drug Discovery; D004789:Enzyme Activation; D020536:Enzyme Activators / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D058851:GPI-Linked Proteins / Q000819:agonists / Q000037:antagonists & inhibitors; D057166:High-Throughput Screening Assays; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors; D051379:Mice; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship
chemical_list: D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D058851:GPI-Linked Proteins; D007527:Isoenzymes; D054852:Small Molecule Libraries; C063088:ALPI protein, human; C548267:ALPL protein, human; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental