Proteomic characterization of the angiogenesis inhibitor SU6668 reveals multiple impacts on cellular kinase signaling.

Knowledge about molecular drug action is critical for the development of protein kinase inhibitors for cancer therapy. Here, we establish a chemical proteomic approach to profile the anticancer drug SU6668, which was originally designed as a selective inhibitor of receptor tyrosine kinases involved in tumor vascularization. By employing immobilized SU6668 for the affinity capture of cellular drug targets in combination with mass spectrometry, we identified previously unknown targets of SU6668 including Aurora kinases and TANK-binding kinase 1. Importantly, a cell cycle block induced by SU6668 could be attributed to inhibition of Aurora kinase activity. Moreover, SU6668 potently suppressed antiviral and inflammatory responses by interfering with TANK-binding kinase 1-mediated signal transmission. These results show the potential of chemical proteomics to provide rationales for the development of potent kinase inhibitors, which combine rather unexpected biological modes of action by simultaneously targeting defined sets of both serine/threonine and tyrosine kinases involved in cancer progression.

[1]  György Kéri,et al.  Cellular targets of gefitinib. , 2005, Cancer research.

[2]  A. Ullrich,et al.  Strategies to overcome resistance to targeted protein kinase inhibitors , 2004, Nature Reviews Drug Discovery.

[3]  M. Cotten,et al.  Chemical Proteomic Analysis Reveals Alternative Modes of Action for Pyrido[2,3-d]pyrimidine Kinase Inhibitors* , 2004, Molecular & Cellular Proteomics.

[4]  Ryan M. O’Connell,et al.  Type I Interferon Production Enhances Susceptibility to Listeria monocytogenes Infection , 2004, The Journal of experimental medicine.

[5]  Michael Karin,et al.  IKKβ Links Inflammation and Tumorigenesis in a Mouse Model of Colitis-Associated Cancer , 2004, Cell.

[6]  G. Müller,et al.  Evaluation of kinase inhibitor selectivity by chemical proteomics. , 2004, Assay and drug development technologies.

[7]  Ola Engkvist,et al.  Characterization of a conserved structural determinant controlling protein kinase sensitivity to selective inhibitors. , 2004, Chemistry & biology.

[8]  J. Ruderman,et al.  Aurora A, Meiosis and Mitosis , 2004, Biology of the cell.

[9]  M. Cotten,et al.  RICK Activates a NF-κB-dependent Anti-human Cytomegalovirus Response* , 2004, Journal of Biological Chemistry.

[10]  David Bebbington,et al.  VX-680, a potent and selective small-molecule inhibitor of the Aurora kinases, suppresses tumor growth in vivo , 2004, Nature Medicine.

[11]  F. Eskens,et al.  Angiogenesis inhibitors in clinical development; where are we now and where are we going? , 2004, British Journal of Cancer.

[12]  T. Maniatis,et al.  IFN-regulatory factor 3-dependent gene expression is defective in Tbk1-deficient mouse embryonic fibroblasts , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. Cotten,et al.  An efficient proteomics method to identify the cellular targets of protein kinase inhibitors , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Swedlow,et al.  Mitotic mechanics: the auroras come into view. , 2003, Current opinion in cell biology.

[15]  D. Hardie.  Printed in U.S.A. Copyright © 2003 by The Endocrine Society doi: 10.1210/en.2003-0982 Minireview: The AMP-Activated Protein Kinase Cascade: The Key Sensor of Cellular Energy Status , 2022 .

[16]  M. Cotten,et al.  Exploiting features of adenovirus replication to support mammalian kinase production. , 2003, Nucleic acids research.

[17]  A. Levitzki.  Protein Kinase Inhibitors as a Therapeutic Modality , 2003 .

[18]  A. Ullrich,et al.  Beyond Herceptin and Gleevec. , 2003, Current opinion in chemical biology.

[19]  F. Mercurio,et al.  IKKi/IKK (cid:1) Plays a Key Role in Integrating Signals Induced by Pro-inflammatory Stimuli* , 2003 .

[20]  Guo-Ping Zhou,et al.  Triggering the Interferon Antiviral Response Through an IKK-Related Pathway , 2003, Science.

[21]  D. Hanahan,et al.  Benefits of targeting both pericytes and endothelial cells in the tumor vasculature with kinase inhibitors. , 2003, The Journal of clinical investigation.

[22]  T. Maniatis,et al.  IKKε and TBK1 are essential components of the IRF3 signaling pathway , 2003, Nature Immunology.

[23]  J. Peters,et al.  The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore–microtubule attachment and in maintaining the spindle assembly checkpoint , 2003, The Journal of cell biology.

[24]  R. Pepperkok,et al.  Chromosome-induced microtubule assembly mediated by TPX2 is required for spindle formation in HeLa cells , 2002, Nature Cell Biology.

[25]  Simon Watkins,et al.  Combined therapy of local and metastatic 4T1 breast tumor in mice using SU6668, an inhibitor of angiogenic receptor tyrosine kinases, and the immunostimulator B7.2-IgG fusion protein. , 2002, Cancer research.

[26]  R. Christopherson,et al.  Inhibitors of de novo nucleotide biosynthesis as drugs. , 2002, Accounts of chemical research.

[27]  J. Cherrington,et al.  SU6668 inhibits Flk‐1/KDR and PDGFRβ in vivo, resulting in rapid apoptosis of tumor vasculature and tumor regression in mice , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[28]  R. Bernards,et al.  A System for Stable Expression of Short Interfering RNAs in Mammalian Cells , 2002, Science.

[29]  J. Cherrington,et al.  Simultaneous inhibition of the receptor kinase activity of vascular endothelial, fibroblast, and platelet-derived growth factors suppresses tumor growth and enhances tumor radiation response. , 2002, Cancer research.

[30]  P. Meraldi,et al.  Aurora‐A overexpression reveals tetraploidization as a major route to centrosome amplification in p53−/− cells , 2002, The EMBO journal.

[31]  C. Allis,et al.  Mitotic Phosphorylation of Histone H3: Spatio-Temporal Regulation by Mammalian Aurora Kinases , 2002, Molecular and Cellular Biology.

[32]  S. Honsawek,et al.  Indolinone tyrosine kinase inhibitors block Kit activation and growth of small cell lung cancer cells. , 2001, Cancer research.

[33]  T. Mak,et al.  Deficiency of T2K leads to apoptotic liver degeneration and impaired NF‐κB‐dependent gene transcription , 2000, The EMBO journal.

[34]  Cherrington,et al.  SU6668 is a potent antiangiogenic and antitumor agent that induces regression of established tumors. , 2000, Cancer research.

[35]  M. Karin,et al.  NAK is an IκB kinase-activating kinase , 2000, Nature.

[36]  Daniel B. Rifkin,et al.  Fibroblast Growth Factor-2 (FGF-2) Induces Vascular Endothelial Growth Factor (VEGF) Expression in the Endothelial Cells of Forming Capillaries: An Autocrine Mechanism Contributing to Angiogenesis , 1998, The Journal of cell biology.

[37]  V. Davisson,et al.  Carboxylases in de novo purine biosynthesis. Characterization of the Gallus gallus bifunctional enzyme. , 1994, Biochemistry.

[38]  Kaori Sasai,et al.  Phosphorylation by aurora kinase A induces Mdm2-mediated destabilization and inhibition of p53 , 2004, Nature Genetics.

[39]  O. Halkka.  [Meiosis and mitosis]. , 1968, Duodecim; laaketieteellinen aikakauskirja.