CNR - Institute of Neuroscience CNR
Institute of Neuroscience
 

Project

Design, characterization and use of kinase inhibitors for signal transduction therapy

Background

Nearly all aspects of cell life and death are controlled by the phosphorylation of proteins which is catalysed by protein kinases (PKs). These belong to the largest family of enzymes, accounting for almost 2% of the proteins encoded by the human genome and collectively referred to as the human "kinome" (Manning et al., 2002). In general PKs are tightly controlled signalling molecules that are switched on only in response to specific stimuli. Not surprisingly therefore the deregulation of PKs results in cell malfunctions, with special reference to neoplastic growth, which makes PKs enticing targets to combat cancer. This gave rise to a neologism, Signal Transduction Therapy (STT), to indicate the specific inhibition of a signalling protein, generally a protein kinase, whose abnormally high activity is causative of neoplasia, as opposed to "chemotherapy" which addresses the problem in a more cursory manner, by blocking DNA replication and thereafter cell proliferation.

According to a recent estimate the human kinome is the first source of pharmacological targets, possibly accounting for more than 20% of the whole "druggable" genome. The majority of these potentially druggable protein kinases ere implicated in neoplastic transformation and hence referred to as the "onco-kinome", a subset of protein kinases whose down-regulation is expected to be beneficial to cancer treatment. Among newcomers in the "onco-kinome", increasing attention is being devoted to CK2, an acronym derived from the misnomer "casein kinase-2", denoting one of the most pleiotropic Ser/Thr protein kinases, whose continuously increasing list of substrates already includes more than 300 proteins, just the tip of an iceberg, considering that it might be responsible for the generation of more than 10% of the whole eukaryotic phosphoproteome. No pathogenic mutations of CK2 are known, however an elevated level of CK2 activity has been invariably observed in a wide variety of tumors as compared to normal tissues.

Objectives

Optimization of cell permeable specific inhibitors for CK2 and CK1 but also for other kinases (e.g. PIM1 and HIPK2), which would also imply the structural analysis of inhibitors in complex with onco-kinase(s). Validation of the therapeutic potential of kinase specific inhibitors in cellular and animal models.

Methods

Through the derivatization of previously known kinase inhibitors new optimal compounds are searched. By virtual screening of large databases of compounds new scaffolds are identified. All the new inhibitors are tested in vitro on purified target kinases and kinetic constants determined while the overall selectivity is tested on batteries of >70 kinases in collaboration with P. Cohens's group in Dundee.

Publications

  • Ruzzene M, Pinna LA (2010) Addiction to protein kinase CK2: a common denominator of diverse cancer cells? Biochim. Biophys. Acta 1804:499-504.
  • Di Maira G, Brustolon F, Pinna LA, Ruzzene M (2009) Dephosphorylation and inactivation of Akt/PKB is counteracted by protein kinase CK2 in HEK 293T cells. Cell. Mol. Life Sci. 66:3363-73.
  • Cozza G, Mazzorana M, Papinutto E, Bain J, Elliott M, di Maira G, Gianoncelli A, Pagano MA, Sarno S, Ruzzene M, Battistutta R, Meggio F, Moro S, Zagotto G, Pinna LA (2009) Quinalizarin as a potent, selective and cell-permeable inhibitor of protein kinase CK2. Biochem. J. 421:387-95.
  • Cozza G, Gianoncelli A, Montopoli M, Caparrotta L, Venerando A, Meggio F, Pinna LA, Zagotto G, Moro S (2008) Identification of novel protein kinase CK1 delta (CK1delta) inhibitors through structure-based virtual screening. Bioorg. Med. Chem. Lett. 18:5672-5.
  • Pagano MA, Bain J, Kazimierczuk Z, Sarno S, Ruzzene M, Di Maira G, Elliott M, Orzeszko A, Cozza G, Meggio F, Pinna LA (2008) The selectivity of inhibitors of protein kinase CK2: an update. Biochem. J. 415:353-65.
  • Di Maira G, Brustolon F, Tosoni K, Belli S, Krämer SD, Pinna LA, Ruzzene M (2008) Comparative analysis of CK2 expression and function in tumor cell lines displaying sensitivity vs. resistance to chemical induced apoptosis. Mol. Cell. Biochem. 316:155-61.
  • Di Maira G, Brustolon F, Bertacchini J, Tosoni K, Marmiroli S, Pinna LA, Ruzzene M (2007) Pharmacological inhibition of protein kinase CK2 reverts the multidrug resistance phenotype of a CEM cell line characterized by high CK2 level. Oncogene 26:6915-26.
  • Pagano MA, Poletto G, Di Maira G, Cozza G, Ruzzene M, Sarno S, Bain J, Elliott M, Moro S, Zagotto G, Meggio F, Pinna LA (2007) Tetrabromocinnamic acid (TBCA) and related compounds represent a new class of specific protein kinase CK2 inhibitors. Chembiochem 8:129-39.
  • Pagano MA, Cesaro L, Meggio F, Pinna LA (2006) Protein kinase CK2: a newcomer in the 'druggable kinome'. Biochem. Soc. Trans. 34:1303-6.

Grants

AIRC

EU, Grant Number: Prokinase research LSHB-CT-2004-503467

Collaborations

  • Prof. Z. Kazimierczuk, Laboratory of Experimental Pharmacology, University of Warsaw, Poland.
  • Prof. G. Zagotto and Prof. S. Moro, Dipartimento di Scienze Farmaceutiche, University of Padova, Italy.

 

PI photo

Lorenzo Pinna

Contact information

email  E-mail

email  049 8276108

Participating staff

Flavio Meggio
Arianna Donella
Roberto Battistutta
Stefania Sarno
Maria Ruzzene
Giovanni Di Maira
Mario A. Pagano
Giorgio Cozza
Elena Papinutto