A chemical genetic method for generating bivalent inhibitors of protein kinases.

We report a new chemical genetic method for creating bivalent ligands of protein kinases. The kinase inhibitors that are generated with this methodology consist of two components: (1) a synthetic, small molecule that targets the ATP-binding cleft and (2) a peptidic ligand that enhances selectivity between kinases by targeting a secondary binding domain. A key feature of these bivalent inhibitors is that they are assembled on a protein scaffold with a chemoselective protein labeling technique. The utility of this methodology is demonstrated through the generation of a panel of protein-small molecule conjugates that simultaneously target the SH1 and SH3 domains of the closely related tyrosine kinases Src and Abl. The assembled bivalent ligands are significantly more potent inhibitors of Src and Abl than either modular component alone. Importantly, these protein-small molecule conjugates show a high degree of selectivity for their intended kinase target.

[1]  Dustin J Maly,et al.  Design, synthesis and characterization of "clickable" 4-anilinoquinazoline kinase inhibitors. , 2008, Molecular bioSystems.

[2]  S. Knapp,et al.  A systematic interaction map of validated kinase inhibitors with Ser/Thr kinases , 2007, Proceedings of the National Academy of Sciences.

[3]  I. Ghosh,et al.  Tethering small molecules to a phage display library: discovery of a selective bivalent inhibitor of protein kinase A. , 2007, Journal of the American Chemical Society.

[4]  K. Shokat,et al.  Chemical Genetics: Where Genetics and Pharmacology Meet , 2007, Cell.

[5]  A. Schepartz,et al.  Encodable activators of SRC family kinases. , 2006, Journal of the American Chemical Society.

[6]  David S Lawrence,et al.  Acquisition of a "Group A"-selective Src kinase inhibitor via a global targeting strategy. , 2006, Journal of the American Chemical Society.

[7]  Oliver Hantschel,et al.  Organization of the SH3-SH2 unit in active and inactive forms of the c-Abl tyrosine kinase. , 2006, Molecular cell.

[8]  D. Schwarzer,et al.  Protein kinase structure and function analysis with chemical tools. , 2005, Biochimica et biophysica acta.

[9]  Kevan M Shokat,et al.  Features of selective kinase inhibitors. , 2005, Chemistry & biology.

[10]  A. Schepartz,et al.  Increasing the kinase specificity of k252a by protein surface recognition. , 2005, Organic letters.

[11]  T. Boggon,et al.  Structure and regulation of Src family kinases , 2004, Oncogene.

[12]  H. Vogel,et al.  Labeling of fusion proteins with synthetic fluorophores in live cells. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[13]  H. Vogel,et al.  Labeling of fusion proteins of O6-alkylguanine-DNA alkyltransferase with small molecules in vivo and in vitro. , 2004, Methods.

[14]  P. Plé,et al.  Discovery of a new class of anilinoquinazoline inhibitors with high affinity and specificity for the tyrosine kinase domain of c-Src. , 2004, Journal of medicinal chemistry.

[15]  P. Cole,et al.  Conversion of a tyrosine kinase protein substrate to a high affinity ligand by ATP linkage. , 2003, Journal of the American Chemical Society.

[16]  N. Johnsson,et al.  Covalent and selective immobilization of fusion proteins. , 2003, Journal of the American Chemical Society.

[17]  D. Lawrence.  Chemical probes of signal-transducing proteins. , 2003, Accounts of chemical research.

[18]  A. Juillerat,et al.  Directed evolution of O6-alkylguanine-DNA alkyltransferase for efficient labeling of fusion proteins with small molecules in vivo. , 2003, Chemistry & biology.

[19]  T. Hunter,et al.  The Protein Kinase Complement of the Human Genome , 2002, Science.

[20]  D. Lawrence,et al.  Molecular Rulers: An Assessment of Distance and Spatial Relationships of Src Tyrosine Kinase SH2 and Active Site Regions* , 2001, The Journal of Biological Chemistry.

[21]  L Serrano,et al.  Rational design of specific high-affinity peptide ligands for the Abl-SH3 domain. , 1996, Biochemistry.

[22]  S. Schreiber,et al.  Specific interactions outside the proline-rich core of two classes of Src homology 3 ligands. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[23]  R. Rickles,et al.  Phage display selection of ligand residues important for Src homology 3 domain binding specificity. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Mindy I. Davis,et al.  A quantitative analysis of kinase inhibitor selectivity , 2008, Nature Biotechnology.

[25]  Sam A. Johnson,et al.  Kinomics: methods for deciphering the kinome , 2004, Nature Methods.

[26]  H. Vogel,et al.  A general method for the covalent labeling of fusion proteins with small molecules in vivo , 2003, Nature Biotechnology.

[27]  S. Hubbard,et al.  Mechanism-based design of a protein kinase inhibitor , 2001, Nature Structural Biology.

[28]  C. Sergheraert,et al.  Design of potent protein kinase inhibitors using the bisubstrate approach. , 1991, Journal of medicinal chemistry.