PHA-680626 Is an Effective Inhibitor of the Interaction between Aurora-A and N-Myc.

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Boi, Dalila 
Souvalidou, Fani 
Polverino, Federica  ORCID logo
Marini, Grazia 

Neuroblastoma is a severe childhood disease, accounting for ~10% of all infant cancers. The amplification of the MYCN gene, coding for the N-Myc transcription factor, is an essential marker correlated with tumor progression and poor prognosis. In neuroblastoma cells, the mitotic kinase Aurora-A (AURKA), also frequently overexpressed in cancer, prevents N-Myc degradation by directly binding to a highly conserved N-Myc region. As a result, elevated levels of N-Myc are observed. During recent years, it has been demonstrated that some ATP competitive inhibitors of AURKA also cause essential conformational changes in the structure of the activation loop of the kinase that prevents N-Myc binding, thus impairing the formation of the AURKA/N-Myc complex. In this study, starting from a screening of crystal structures of AURKA in complexes with known inhibitors, we identified additional compounds affecting the conformation of the kinase activation loop. We assessed the ability of such compounds to disrupt the interaction between AURKA and N-Myc in vitro, using Surface Plasmon Resonance competition assays, and in tumor cell lines overexpressing MYCN, by performing Proximity Ligation Assays. Finally, their effects on N-Myc cellular levels and cell viability were investigated. Our results identify PHA-680626 as an amphosteric inhibitor both in vitro and in MYCN overexpressing cell lines, thus expanding the repertoire of known conformational disrupting inhibitors of the AURKA/N-Myc complex and confirming that altering the conformation of the activation loop of AURKA with a small molecule is an effective strategy to destabilize the AURKA/N-Myc interaction in neuroblastoma cancer cells.

Aurora-A, N-Myc, PHA-680626, amphosteric inhibitors, neuroblastoma, Adenosine Triphosphate, Antineoplastic Agents, Aurora Kinase A, Azepines, Benzazepines, Binding Sites, Binding, Competitive, Cell Line, Drug Evaluation, Preclinical, Humans, N-Myc Proto-Oncogene Protein, Neuroblastoma, Protein Conformation, Protein Kinase Inhibitors, Pyrazoles, Pyrimidines, Pyrroles, Surface Plasmon Resonance
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Int J Mol Sci
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Italian Association for Cancer Research (MFAG 20447)