(S)‐Pyrrolo[1,2‐a]pyrazine‐3‐carboxamide Analogs as Dual Akt/JNK Modulators With Antiproliferative Effects in MDA‐MB‐468 Cells

Abstract

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.