Anticancer Activity of (S)-5-Chloro-3-((3,5-dimethylphenyl)sulfonyl)- N-(1-oxo-1-((pyridin-4-ylmethyl)amino)propan-2-yl)-1 H-indole-2-carboxamide (RS4690), a New Dishevelled 1 Inhibitor

Antonio Coluccia; Marianna Bufano; Giuseppe La Regina; Michela Puxeddu; Angelo Toto; Alessio Paone; Amani Bouzidi; Giorgia Musto; Nadia Badolati; Viviana Orlando; Stefano Biagioni; Domiziana Masci; Chiara Cantatore; Roberto Cirilli; Francesca Cutruzzolà; Stefano Gianni; Mariano Stornaiuolo; Romano Silvestri

doi:10.3390/cancers14051358

Anticancer Activity of (S)-5-Chloro-3-((3,5-dimethylphenyl)sulfonyl)- N-(1-oxo-1-((pyridin-4-ylmethyl)amino)propan-2-yl)-1 H-indole-2-carboxamide (RS4690), a New Dishevelled 1 Inhibitor

Cancers (Basel). 2022 Mar 7;14(5):1358. doi: 10.3390/cancers14051358.

Authors

Antonio Coluccia¹, Marianna Bufano¹, Giuseppe La Regina¹, Michela Puxeddu¹, Angelo Toto², Alessio Paone², Amani Bouzidi², Giorgia Musto³, Nadia Badolati³, Viviana Orlando⁴, Stefano Biagioni⁴, Domiziana Masci⁵, Chiara Cantatore⁶, Roberto Cirilli⁶, Francesca Cutruzzolà², Stefano Gianni², Mariano Stornaiuolo³, Romano Silvestri¹

Affiliations

¹ Laboratory Affiliated with the Institute Pasteur Italy-Cenci Bolognetti Foundation, Department of Drug Chemistry and Technologies, Sapienza University of Rome, Piazzale Aldo Moro 5, 00185 Rome, Italy.
² Laboratory Affiliated with the Institute Pasteur Italy-Cenci Bolognetti Foundation, Biochemical Sciences "Rossi Fanelli", Institute of Biology and Molecular Pathology of CNR, Sapienza Università di Roma, Piazzale Aldo Moro 5, 00185 Rome, Italy.
³ Department of Pharmacy, University of Naples "Federico II", Via Domenico Montesano, 80131 Naples, Italy.
⁴ Department of Biology and Biotechnologies "Charles Darwin", Sapienza University of Rome, Piazzale Aldo Moro 5, 00185 Roma, Italy.
⁵ Department of Basic Biotechnological Sciences, Intensivological and Perioperative Clinics, Catholic University of Sacred Heart, Largo Francesco Vito 1, 00168 Rome, Italy.
⁶ National Center for the Control and Evaluation of Drugs, Istituto Superiore di Sanità, Rome, Viale Regina Elena 299, 00161 Rome, Italy.

Abstract

Wingless/integrase-11 (WNT)/β-catenin pathway is a crucial upstream regulator of a huge array of cellular functions. Its dysregulation is correlated to neoplastic cellular transition and cancer proliferation. Members of the Dishevelled (DVL) family of proteins play an important role in the transduction of WNT signaling by contacting its cognate receptor, Frizzled, via a shared PDZ domain. Thus, negative modulators of DVL1 are able to impair the binding to Frizzled receptors, turning off the aberrant activation of the WNT pathway and leading to anti-cancer activity. Through structure-based virtual screening studies, we identified racemic compound RS4690 (1), which showed a promising selective DVL1 binding inhibition with an EC₅₀ of 0.74 ± 0.08 μM. Molecular dynamic simulations suggested a different binding mode for the enantiomers. In the in vitro assays, enantiomer (S)-1 showed better inhibition of DVL1 with an EC₅₀ of 0.49 ± 0.11 μM compared to the (R)-enantiomer. Compound (S)-1 inhibited the growth of HCT116 cells expressing wild-type APC with an EC₅₀ of 7.1 ± 0.6 μM and caused a high level of ROS production. These results highlight (S)-1 as a lead compound for the development of new therapeutic agents against WNT-dependent colon cancer.

Keywords: DVL1; PDZ domain; WNT pathway; cancer; virtual screening.