title: Mechanism and in vitro pharmacology of TAK1 inhibition by (5Z)-7-Oxozeaenol.
abstract: Transforming growth factor-β activated kinase-1 (TAK1) is a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family that regulates several signaling pathways including NF-κB signal transduction and p38 activation. TAK1 deregulation has been implicated in human diseases including cancer and inflammation. Here, we show that, in addition to its kinase activity, TAK1 has intrinsic ATPase activity, that (5Z)-7-Oxozeaenol irreversibly inhibits TAK1, and that sensitivity to (5Z)-7-Oxozeaenol inhibition in hematological cancer cell lines is NRAS mutation status and TAK1 pathway dependent. X-ray crystallographic and mass spectrometric studies showed that (5Z)-7-Oxozeaenol forms a covalent complex with TAK1. Detailed biochemical characterization revealed that (5Z)-7-Oxozeaenol inhibited both the kinase and the ATPase activity of TAK1 following a bi-phase kinetics, consistent with the irreversible inhibition mechanism. In DoHH2 cells, (5Z)-7-Oxozeaenol potently inhibited the p38 phosphorylation driven by TAK1, and the inhibition lasted over 6 h after withdrawal of (5Z)-7-Oxozeaenol. Profiling (5Z)-7-Oxozeaenol in a panel of hematological cancer cells showed that sensitive cell lines tended to carry NRAS mutations and that genes in TAK1 regulated pathways were enriched in sensitive cell lines. Taken together, we have elucidated the molecular mechanism of a TAK1 irreversible inhibitor and laid the foundation for designing next generation TAK1 irreversible inhibitors. The NRAS-TAK1-Wnt signaling network discerned in our study may prove to be useful in patient selection for TAK1 targeted agents in hematological cancers.
authors.affiliation: Discovery Sciences, AstraZeneca R&D Boston, Waltham, MA 02451, USA.
authors.forename: Jiaquan
authors.initials: J
authors.lastname: Wu
authors.forename: Francoise
authors.initials: F
authors.lastname: Powell
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Larsen
authors.forename: Zhongwu
authors.initials: Z
authors.lastname: Lai
authors.forename: Kate F
authors.initials: KF
authors.lastname: Byth
authors.forename: Jon
authors.initials: J
authors.lastname: Read
authors.forename: Rong-Fang
authors.initials: RF
authors.lastname: Gu
authors.forename: Mark
authors.initials: M
authors.lastname: Roth
authors.forename: Dorin
authors.initials: D
authors.lastname: Toader
authors.forename: Jamal Carlos
authors.initials: JC
authors.lastname: Saeh
authors.forename: Huawei
authors.initials: H
authors.lastname: Chen
mesh_terms: D045744:Cell Line, Tumor; D018360:Crystallography, X-Ray; D004305:Dose-Response Relationship, Drug; D006801:Humans; D020930:MAP Kinase Kinase Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D015025:Zearalenone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology / Q000008:administration & dosage; D000099151:MAP Kinase Kinase Kinase 7; D007783:Lactones; D012118:Resorcinols
chemical_list: D020930:MAP Kinase Kinase Kinases; D047428:Protein Kinase Inhibitors; D015025:Zearalenone; D000099151:MAP Kinase Kinase Kinase 7; C505734:7-oxozeanol; D007783:Lactones; D012118:Resorcinols