BRAF is a serine/threonine kinase frequently mutated in human cancers. BRAFV600E mutated protein is targeted through the use of kinase inhibitors which are approved for the treatment of melanoma; however, their long-term efficacy is hampered by resistance mechanisms. The PROTAC-induced degradation of BRAFV600E has been proposed as an alternative strategy to avoid the onset of resistance. In this study, we designed a series of compounds where the BRAF kinase inhibitor encorafenib was conjugated to pomalidomide through different linkers. The synthesized compounds maintained their ability to inhibit the kinase activity of mutated BRAF with IC50 values in the 40-88 nM range. Selected compounds inhibited BRAFV600E signaling and cellular proliferation of A375 and Colo205 tumor cell lines. Compounds 10 and 11, the most active of the series, were not able to induce degradation of mutated BRAF. Docking and molecular dynamic studies, conducted in comparison with the efficient BRAF degrader P5B, suggest that a different orientation of the linker bearing the pomalidomide substructure, together with a decreased mobility of the solvent-exposed part of the conjugates, could explain this behavior.

Investigation into the Use of Encorafenib to Develop Potential PROTACs Directed against BRAFV600E Protein

Marini, Elisabetta;Marino, Marco;Gionfriddo, Giulia;Maione, Federica;Oddo, Daniele;Giorgis, Marta;Rolando, Barbara;Blua, Federica;Gastaldi, Simone;Marchiò, Serena;Kovachka, Sandra;Spyrakis, Francesca;Gianquinto, Eleonora
;
Di Nicolantonio, Federica
;
Bertinaria, Massimo
2022-01-01

Abstract

BRAF is a serine/threonine kinase frequently mutated in human cancers. BRAFV600E mutated protein is targeted through the use of kinase inhibitors which are approved for the treatment of melanoma; however, their long-term efficacy is hampered by resistance mechanisms. The PROTAC-induced degradation of BRAFV600E has been proposed as an alternative strategy to avoid the onset of resistance. In this study, we designed a series of compounds where the BRAF kinase inhibitor encorafenib was conjugated to pomalidomide through different linkers. The synthesized compounds maintained their ability to inhibit the kinase activity of mutated BRAF with IC50 values in the 40-88 nM range. Selected compounds inhibited BRAFV600E signaling and cellular proliferation of A375 and Colo205 tumor cell lines. Compounds 10 and 11, the most active of the series, were not able to induce degradation of mutated BRAF. Docking and molecular dynamic studies, conducted in comparison with the efficient BRAF degrader P5B, suggest that a different orientation of the linker bearing the pomalidomide substructure, together with a decreased mobility of the solvent-exposed part of the conjugates, could explain this behavior.
2022
27
23
8513
8546
BRAF kinase; PROTACs; encorafenib; molecular dynamics; pomalidomide
Marini, Elisabetta; Marino, Marco; Gionfriddo, Giulia; Maione, Federica; Pandini, Marta; Oddo, Daniele; Giorgis, Marta; Rolando, Barbara; Blua, Federica; Gastaldi, Simone; Marchiò, Serena; Kovachka, Sandra; Spyrakis, Francesca; Gianquinto, Eleonora; Di Nicolantonio, Federica; Bertinaria, Massimo
File in questo prodotto:
File Dimensione Formato  
molecules-27-08513-v2.pdf

Accesso aperto

Tipo di file: PDF EDITORIALE
Dimensione 7.88 MB
Formato Adobe PDF
7.88 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/1882728
Citazioni
  • ???jsp.display-item.citation.pmc??? 3
  • Scopus 5
  • ???jsp.display-item.citation.isi??? 5
social impact