@Article{or.2022.025187,
AUTHOR = {FAISAL HASSAN TOBEIGEI, REEM M. GAHTANI, AHMAD SHAIKH, AMER AL ALI, NADER KAMELI, HOSSAM KAMLI, PRASANNA RAJAGOPALAN},
TITLE = {Computational high-throughput screening and <i>in vitro</i> approaches identify CB-006-3; A novel PI3K-BRAF<sup>V600E</sup> dual targeted inhibitor against melanoma},
JOURNAL = {Oncology Research},
VOLUME = {29},
YEAR = {2021},
NUMBER = {5},
PAGES = {305--318},
URL = {http://www.techscience.com/or/v29n5/50095},
ISSN = {1555-3906},
ABSTRACT = {Malignant melanoma is characterized by both genetic and molecular alterations that activate phosphoinositide 3-kinase (PI3K), and RAS/BRAF pathways. In this work, through diversity-based high-throughput virtual screening we identified a lead molecule that selectively targets PI3K and BRAF<sup>V600E</sup> kinases. Computational screening, Molecular dynamics simulation and MMPBSA calculations were performed. PI3K and BRAF<sup>V600E</sup> kinase inhibition was done. A375 and G-361 cells were used for <i>in vitro</i> cellular analysis to determine antiproliferative effects, annexin V binding, nuclear fragmentation and cell cycle analysis. Computational screening of small molecules indicates compound CB-006-3 selectively targets PI3KCG (gamma subunit), PI3KCD (delta subunit) and BRAF<sup>V600E</sup>. Molecular dynamics simulation and MMPBSA bases binding free energy calculations predict a stable binding of CB-006-3 to the active sites of PI3K and BRAF<sup>V600E</sup>. The compound effectively inhibited PI3KCG, PI3KCD and BRAF<sup>V600E</sup> kinases with respective IC<sub>50</sub> values of 75.80, 160.10 and 70.84 nM. CB-006-3 controlled the proliferation of A375 and G-361 cells with GI<sub>50</sub> values of 223.3 and 143.6 nM, respectively. A dose dependent increase in apoptotic cell population and sub G<sub>0</sub>/G<sub>1</sub> phase of cell cycle were also observed with the compound treatment in addition to observed nuclear fragmentation in these cells. Furthermore, CB-006-3 inhibited BRAF<sup>V600E</sup>, PI3KCD and PI3KCG in both melanoma cells. Collectively, based on the computational modeling and <i>in vitro</i> validations, we propose CB-006-3 as a lead candidate for selectively targeting PI3K and mutant BRAF<sup>V600E</sup> to inhibit melanoma cell proliferation. Further experimental validations, including pharmacokinetic evaluations in mouse models will identify the druggability of the proposed lead candidate for further development as a therapeutic agent for treating melanoma.},
DOI = {10.32604/or.2022.025187}
}