Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61

• Main Authors: Andrea Calcaterra (Author), Valentina Iovine (Author), Bruno Botta (Author), Deborah Quaglio (Author), Ilaria D'Acquarica (Author), Alessia Ciogli (Author), Antonia Iazzetti (Author), Romina Alfonsi (Author), Ludovica Lospinoso Severini (Author), Paola Infante (Author), Lucia Di Marcotullio (Author), Mattia Mori (Author), Francesca Ghirga (Author)
• Format: Book
• Published: Taylor & Francis Group, 2018-01-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

This work aims at elucidating the mechanism and kinetics of hydrolysis of GANT61, the first and most-widely used inhibitor of the Hedgehog (Hh) signalling pathway that targets Glioma-associated oncogene homologue (Gli) proteins, and at confirming the chemical nature of its bioactive form. GANT61 is poorly stable under physiological conditions and rapidly hydrolyses into an aldehyde species (GANT61-A), which is devoid of the biological activity against Hh signalling, and a diamine derivative (GANT61-D), which has shown inhibition of Gli-mediated transcription. Here, we combined chemical synthesis, NMR spectroscopy, analytical studies, molecular modelling and functional cell assays to characterise the GANT61 hydrolysis pathway. Our results show that GANT61-D is the bioactive form of GANT61 in NIH3T3 Shh-Light II cells and SuFu−/− mouse embryonic fibroblasts, and clarify the structural requirements for GANT61-D binding to Gli1. This study paves the way to the design of GANT61 derivatives with improved potency and chemical stability.

Item Description

1475-6366
1475-6374
10.1080/14756366.2017.1419221