title: Damnacanthal is a highly potent, selective inhibitor of p56lck tyrosine kinase activity.
abstract: Damnacanthal, an anthraquinone isolated from a plant extract, was found to be a potent, selective inhibitor of p56lck tyrosine kinase activity. The structure, potency, and selectivity of damnacanthal were confirmed by independent synthesis and testing. Damnacanthal exhibited an IC50 of 17 nM for inhibition of p56lck autophosphorylation and an IC50 of 620 nM for phosphorylation of an exogenous peptide by p56lck. Damnacanthal had > 100-fold selectivity for p56lck over the serine/threonine kinases, protein kinase A and protein kinase C, and > 40-fold selectivity for p56lck over four receptor tyrosine kinases. It also demonstrated modest (7-20-fold), but highly statistically significant, selectivity for p56lck over the homologous enzymes p60src and p59fyn. Mechanistic studies demonstrated that damnacanthal was competitive with the peptide binding site, but mixed noncompetitive with the ATP site. Although damnacanthal contains a potentially reactive aldehyde moiety, equilibrium dialysis experiments demonstrated that significant amine formation between damnacanthal and amines occurred only at high concentrations of reactants. However, damnacanthal appeared to bind nonspecifically to membrane lipids and was not active in whole cell tyrosine kinase assays. Damnacanthal is the most potent, selective inhibitor of p56lck tyrosine kinase activity described to date and may represent the starting point for the identification of novel, selective inhibitors of p56lck which are active in whole cell as well as in cell-free systems.
authors.affiliation: Department of Immunology, Sterling Winthrop Pharmaceuticals Research Division, Collegeville, Pennsylvania 19426, USA.
authors.forename: C R
authors.initials: CR
authors.lastname: Faltynek
authors.forename: J
authors.initials: J
authors.lastname: Schroeder
authors.forename: P
authors.initials: P
authors.lastname: Mauvais
authors.forename: D
authors.initials: D
authors.lastname: Miller
authors.forename: S
authors.initials: S
authors.lastname: Wang
authors.forename: D
authors.initials: D
authors.lastname: Murphy
authors.forename: R
authors.initials: R
authors.lastname: Lehr
authors.forename: M
authors.initials: M
authors.lastname: Kelley
authors.forename: A
authors.initials: A
authors.lastname: Maycock
authors.forename: W
authors.initials: W
authors.lastname: Michne
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D000880:Anthraquinones / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007700:Kinetics; D008055:Lipids / Q000494:pharmacology; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011107:Polylysine / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000187:drug effects; D011505:Protein-Tyrosine Kinases / Q000187:drug effects; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D019311:Staurosporine; D019061:src-Family Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000470:Alkaloids; D000880:Anthraquinones; D004791:Enzyme Inhibitors; D008055:Lipids; D010455:Peptides; D011107:Polylysine; C079170:damnacanthal; D011505:Protein-Tyrosine Kinases; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D019061:src-Family Kinases; D017346:Protein Serine-Threonine Kinases; D012259:Ribonuclease, Pancreatic; D019311:Staurosporine