Pyrimidine-5-carbonitrile based potential anticancer agents as apoptosis inducers through PI3K/AKT axis inhibition in leukaemia K562

Abstract

A novel series of 4-(4-Methoxyphenyl)-2-(methylthio)pyrimidine-5-carbonitrile was developed linked to an aromatic moiety <i>via N</i>-containing bridge and then evaluated for their cytotoxic activity against MCF-7 and K562 cell lines. Seven compounds exhibited the highest activity against both cell lines where compounds <b>4d</b> and <b>7f</b> were the most active against K562 cell line. Exploring their molecular mechanisms by enzyme inhibition assay on PI3Kδ/γ and AKT-1 showed that compound <b>7f</b> was promising more than <b>4d</b> with IC₅₀ = 6.99 ± 0.36, 4.01 ± 0.55, and 3.36 ± 0.17 uM, respectively. Also, flowcytometric analysis revealed that <b>7f</b> caused cell cycle arrest at S-phase followed by caspase 3 dependent apoptosis induction. Mechanistically, compound <b>7f</b> proved to modulate the expression of PI3K, p-PI3K, AKT, p-AKT, Cyclin D1, and NFΚβ. Furthermore, <i>in-vivo</i> toxicity study indicated good safety profile for <b>7f</b>. These findings suggest that the trimethoxy derivative <b>7f</b> has strong potential as a multi-acting inhibitor on PI3K/AKT axis targeting breast cancer and leukaemia.