(S)-Pyrrolo[1,2-a]pyrazine-3-carboxamide Analogs as Dual Akt/JNK Modulators With Antiproliferative Effects in...

Triple-negative breast cancer (TNBC) remains a major clinical challenge due to the absence of effective targeted therapies. In this study, thirty analogs of (S)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-3-carboxamide (A1-F5) were designed, synthesized, and evaluated for their cytotoxic activity against hormone receptor-positive breast cancer (MCF-7) and EGFR-overexpressing TNBC (MDA-MB-468) cell lines. Notably, compounds B1 and B3 exhibited potent growth inhibition toward MDA-MB-468 cells, with GI₅₀ values of 2.8 and 4.7 µM, respectively, surpassing the efficacy of the reference compound (Ref 2, GI₅₀: 7.6 µM) and gefitinib (GI₅₀: 16.5 µM). Flow cytometric analysis demonstrated significant apoptosis induction by compounds B1 and B3, with rates of 42.8% and 42.2%, respectively. Mechanistic investigations revealed that compounds B1 and B3 selectively inhibited Akt phosphorylation and enhanced JNK phosphorylation, suggesting a dual modulation of survival and apoptotic pathways. These findings support the development of N2-benzyl-(S)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-3-carboxamide analogs, particularly compound B1, as selective therapeutic candidates for triple-negative breast cancer.