The antisignaling agent SC-alpha alpha delta 9, 4-(benzyl-(2-[(2,5-diphenyloxazole-4-carbonyl)amino]ethyl)carbamoyl)- 2-decanoylaminobutyric acid, is a structurally unique phospholipid analogue with phospholipase C inhibitory activity.
Phospholipids and lipid second messengers mediate mitogenic signal transduction and oncogenesis, but there have been few successful examples of small molecules that affect biologically important phospholipid metabolism. Here we investigated the actions of a previously described antitumor agent, 4-(benzyl-(2-[(2,5-diphenyloxazole-4-carbonyl)amino]ethyl)carbamoyl)- 2-decanoylaminobutyric acid (SC-alpha alpha delta 9), which has antisignaling properties, on phospholipases. Although SC-alpha alpha delta 9 had been shown to be a potent and selective inhibitor of the Cdc25 family of dual-specificity phosphatases, many of its cellular effects are not readily reconciled with phosphatase inhibition. Molecular modeling studies suggested that SC-alpha alpha delta 9 shared several structural features with membrane phospholipids. Enzyme inhibition studies in vitro revealed that SC-alpha alpha delta 9 was a potent inhibitor of phospholipase C (PLC; IC50 = 25 microM) but did not inhibit phospholipase D activity at concentrations up to 100 microM. In H-ras (Q61L)-transformed Rat-1 fibroblasts with constitutively elevated levels of phosphorylated extracellular signal-regulated kinase (Erk), SC-alpha alpha delta 9 inhibited both proliferation and oncogenic Erk activation at concentrations that inhibited PLC in vitro. A SC-alpha alpha delta 9 congener that lacked antiproliferative activity also did not inhibit PLC in vitro. In the PLC-dependent scratch wound healing model, SC-alpha alpha delta 9 was 10-fold more potent than the phosphatidylcholine-specific PLC inhibitor D-609. We propose that the structural resemblance of SC-alpha alpha delta 9 to phospholipids allows it to inhibit cellular PLC, thereby providing a possible molecular mechanism for SC-alpha alpha delta 9's effects on oncogenic Erk activation.
Vogt
Andreas
A
Department of Pharmacology, Biomedical Science Tower E-1017, University of Pittsburgh, Pittsburgh, PA 15261, USA. avogt@pitt.edu
Pestell
Katharine E
KE
Day
Billy W
BW
Lazo
John S
JS
Wipf
Peter
P
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D001952:Bridged-Ring Compounds / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D011905:Genes, ras / Q000235:genetics; D020128:Inhibitory Concentration 50; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D009636:Norbornanes; D010080:Oxazoles / Q000737:chemistry / Q000494:pharmacology*; D010739:Phospholipase D / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D013859:Thiocarbamates; D013871:Thiones / Q000494:pharmacology; D013997:Time Factors; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects
D001952:Bridged-Ring Compounds; D004791:Enzyme Inhibitors; D009636:Norbornanes; D010080:Oxazoles; D010743:Phospholipids; C115707:SC-alphaalphadelta9; D013859:Thiocarbamates; D013871:Thiones; C046498:tricyclodecane-9-yl-xanthogenate; D020928:Mitogen-Activated Protein Kinases; D000469:Alkaline Phosphatase; D010738:Type C Phospholipases; D010739:Phospholipase D