title: Potent and selective disruption of protein kinase D functionality by a benzoxoloazepinolone.
abstract: Protein kinase D (PKD) is a novel family of serine/threonine kinases targeted by the second messenger diacylglycerol. It has been implicated in many important cellular processes and pathological conditions. However, further analysis of PKD in these processes is severely hampered by the lack of a PKD-specific inhibitor that can be readily applied to cells and in animal models. We now report the discovery of the first potent and selective cell-active small molecule inhibitor for PKD, benzoxoloazepinolone (CID755673). This inhibitor was identified from the National Institutes of Health small molecule repository library of 196,173 compounds using a human PKD1 (PKCmu)-based fluorescence polarization high throughput screening assay. CID755673 suppressed half of the PKD1 enzyme activity at 182 nm and exhibited selective PKD1 inhibition when compared with AKT, polo-like kinase 1 (PLK1), CDK activating kinase (CAK), CAMKIIalpha, and three different PKC isoforms. Moreover, it was not competitive with ATP for enzyme inhibition. In cell-based assays, CID755673 blocked phorbol ester-induced endogenous PKD1 activation in LNCaP cells in a concentration-dependent manner. Functionally, CID755673 inhibited the known biological actions of PKD1 including phorbol ester-induced class IIa histone deacetylase 5 nuclear exclusion, vesicular stomatitis virus glycoprotein transport from the Golgi to the plasma membrane, and the ilimaquinone-induced Golgi fragmentation. Moreover, CID755673 inhibited prostate cancer cell proliferation, cell migration, and invasion. In summary, our findings indicate that CID755673 is a potent and selective PKD1 inhibitor with valuable pharmacological and cell biological potential.
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Sharlow
authors.forename: Karthik V
authors.initials: KV
authors.lastname: Giridhar
authors.forename: Courtney R
authors.initials: CR
authors.lastname: LaValle
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
authors.forename: Stephanie
authors.initials: S
authors.lastname: Leimgruber
authors.forename: Rebecca
authors.initials: R
authors.lastname: Barrett
authors.forename: Karla
authors.initials: K
authors.lastname: Bravo-Altamirano
authors.forename: Peter
authors.initials: P
authors.lastname: Wipf
authors.forename: John S
authors.initials: JS
authors.lastname: Lazo
authors.forename: Q Jane
authors.initials: QJ
authors.lastname: Wang
mesh_terms: D001381:Azepines / Q000737:chemistry / Q000494:pharmacology*; D001572:Benzofurans / Q000737:chemistry / Q000494:pharmacology*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D000070819:Discoidin Domain Receptor 1; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D006056:Golgi Apparatus / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D010703:Phorbol Esters / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D000097869:Polo-Like Kinase 1
chemical_list: D001381:Azepines; D001572:Benzofurans; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D018797:Cell Cycle Proteins; D000070819:Discoidin Domain Receptor 1; D010703:Phorbol Esters; D011493:Protein Kinase C; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D020794:Receptor Protein-Tyrosine Kinases; D000097869:Polo-Like Kinase 1; C534615:CID755673; C089212:protein kinase D; C092030:DDR1 protein, human