The commonly used PI3-kinase probe LY294002 is an inhibitor of BET bromodomains.

A commonly used small-molecule probe in cell-signaling research is the phosphoinositide 3-kinase inhibitor LY294002. Quantitative chemoproteomic profiling shows that LY294002 and LY303511, a close analogue devoid of PI3K activity, inhibit the BET bromodomain proteins BRD2, BRD3, and BRD4 that comprise a family of targets structurally unrelated to PI3K. Both compounds competitively inhibit acetyl-lysine binding of the first but not the second bromodomain of BET proteins in cell extracts. X-ray crystallography shows that the chromen-4-one scaffold represents a new bromodomain pharmacophore and establishes LY294002 as a dual kinase and BET-bromodomain inhibitor, whereas LY303511 exhibits anti-inflammatory and antiproliferative effects similar to the recently discovered BET inhibitors.

[1]  S. Olesen,et al.  Cyclin-dependent kinase inhibitor dinaciclib interacts with the acetyl-lysine recognition site of bromodomains. , 2013, ACS chemical biology.

[2]  Mark E Bunnage,et al.  Target validation using chemical probes. , 2013, Nature chemical biology.

[3]  A. Belkina,et al.  BET Protein Function Is Required for Inflammation: Brd2 Genetic Disruption and BET Inhibitor JQ1 Impair Mouse Macrophage Inflammatory Responses , 2013, The Journal of Immunology.

[4]  Jacob K. White,et al.  Reactive oxygen species (ROS) and sensitization to TRAIL-induced apoptosis, in Bayesian network modelling of HeLa cell response to LY303511. , 2012, Biochemical pharmacology.

[5]  J. Greenbaum,et al.  Selective inhibition of CD4+ T-cell cytokine production and autoimmunity by BET protein and c-Myc inhibitors , 2012, Proceedings of the National Academy of Sciences.

[6]  G. Drewes,et al.  Corrigendum: A selective inhibitor reveals PI3Kγ dependence of T H 17 cell differentiation , 2012 .

[7]  G. Drewes,et al.  A Selective Inhibitor Reveals Pi3Kgamma Dependence of T(H)17 Cell Differentiation. , 2012 .

[8]  B. N. Devaiah,et al.  BRD4 is an atypical kinase that phosphorylates Serine2 of the RNA Polymerase II carboxy-terminal domain , 2012, Proceedings of the National Academy of Sciences.

[9]  Jianni Qi,et al.  LY294002 inhibits TLR3/4‐mediated IFN‐β production via inhibition of IRF3 activation with a PI3K‐independent mechanism , 2012, FEBS letters.

[10]  Marcus Bantscheff,et al.  Chemoproteomic approaches to drug target identification and drug profiling. , 2012, Bioorganic & medicinal chemistry.

[11]  Raymond E Moellering,et al.  How chemoproteomics can enable drug discovery and development. , 2012, Chemistry & biology.

[12]  S. Lowe,et al.  RNAi screen identifies Brd4 as a therapeutic target in acute myeloid leukaemia , 2011, Nature.

[13]  S. Robson,et al.  Inhibition of BET recruitment to chromatin as an effective treatment for MLL-fusion leukaemia , 2011, Nature.

[14]  P. Bamborough,et al.  Discovery and characterization of small molecule inhibitors of the BET family bromodomains. , 2011, Journal of medicinal chemistry.

[15]  Jennifer A. Smith,et al.  The Brd4 Extraterminal Domain Confers Transcription Activation Independent of pTEFb by Recruiting Multiple Proteins, Including NSD3 , 2011, Molecular and Cellular Biology.

[16]  P. Grandi,et al.  Chemoproteomics profiling of HDAC inhibitors reveals selective targeting of HDAC complexes , 2011, Nature Biotechnology.

[17]  C. Rice,et al.  Suppression of inflammation by a synthetic histone mimic , 2010, Nature.

[18]  William B. Smith,et al.  Selective inhibition of BET bromodomains , 2010, Nature.

[19]  B. Vanhaesebroeck,et al.  The emerging mechanisms of isoform-specific PI3K signalling , 2010, Nature Reviews Molecular Cell Biology.

[20]  Paul Workman,et al.  Drugging the PI3 kinome: from chemical tools to drugs in the clinic. , 2010, Cancer research.

[21]  K. Ozato,et al.  Brd4 marks select genes on mitotic chromatin and directs postmitotic transcription. , 2009, Molecular biology of the cell.

[22]  J. Cox,et al.  Proteomics strategy for quantitative protein interaction profiling in cell extracts , 2009, Nature Methods.

[23]  G. Superti-Furga,et al.  Target profiling of small molecules by chemical proteomics. , 2009, Nature chemical biology.

[24]  T. Willson,et al.  Open access chemical and clinical probes to support drug discovery. , 2009, Nature chemical biology.

[25]  Bernhard Kuster,et al.  Quantitative chemical proteomics reveals mechanisms of action of clinical ABL kinase inhibitors , 2007, Nature Biotechnology.

[26]  M. Zvelebil,et al.  Exploring the specificity of the PI3K family inhibitor LY294002. , 2007, The Biochemical journal.

[27]  M. Waterfield,et al.  Synthesis and biological evaluation of pyrido[3′,2′:4,5]furo[3,2- d ]pyrimidine derivatives as novel PI3 kinase p110α inhibitors , 2007 .

[28]  P. Hawkins,et al.  Signalling through Class I PI3Ks in mammalian cells. , 2006, Biochemical Society transactions.

[29]  A. Fliri,et al.  Analysis of drug-induced effect patterns to link structure and side effects of medicines , 2005, Nature chemical biology.

[30]  J. Moss,et al.  LY303511 (2-Piperazinyl-8-phenyl-4H-1-benzopyran-4-one) Acts via Phosphatidylinositol 3-Kinase-Independent Pathways to Inhibit Cell Proliferation via Mammalian Target of Rapamycin (mTOR)- and Non-mTOR-Dependent Mechanisms , 2005, Journal of Pharmacology and Experimental Therapeutics.

[31]  T. Kwon,et al.  LY294002 inhibits LPS-induced NO production through a inhibition of NF-kappaB activation: independent mechanism of phosphatidylinositol 3-kinase. , 2005, Immunology letters.

[32]  Anthony C. Bishop,et al.  Rapid structure-activity and selectivity analysis of kinase inhibitors by BioMAP analysis in complex human primary cell-based models. , 2004, Assay and drug development technologies.

[33]  E. Berg,et al.  An integrative biology approach for analysis of drug action in models of human vascular inflammation , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[34]  D. Faller,et al.  You bet-cha: a novel family of transcriptional regulators. , 2001, Frontiers in bioscience : a journal and virtual library.

[35]  Roger L. Williams,et al.  Structural determinants of phosphoinositide 3-kinase inhibition by wortmannin, LY294002, quercetin, myricetin, and staurosporine. , 2000, Molecular cell.

[36]  M. Green,et al.  A novel, mitogen-activated nuclear kinase is related to a Drosophila developmental regulator. , 1996, Genes & development.

[37]  K Y Hui,et al.  A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). , 1994, The Journal of biological chemistry.