Design, Synthesis and Evaluation of Anticancer Potential of Novel Pyrrolopyrazines Against Glioblastoma: Mechanistic Details and In Vivo Safety Analysis

A protocol through propargyl moiety on pyrrole ring cyclization with ammonium acetate was demonstrated. With this protocol, 11 pyrrolo1,2-apyrazine derivatives were synthesized, and their antiproliferative activity against the U87MG glioblastoma cell line was evaluated. Among the synthesized compounds, a naphthyl-substituted derivative (3g) exhibited the most potent in vitro activity with an IC50 value of 60 µM, and showed no antiproliferative effect against human dermal fibroblast (HDF) cells up to 100 μM. Gene expression analysis of U87MG cells treated with 3g revealed a significant upregulation of key genes involved in apoptosis (CASP8, CASP9, CASP3, BAX, BCL2), autophagy (ATG3, ATG5), necroptosis (RIPK1), and DNA damage repair (PARP1) suggesting a multifaceted cell death mechanism. Furthermore, the in vivo safety profile of compound 3g was assessed in an animal model by evaluating renal (BUN, creatinine) and hepatic (AST, ALT, ALP) biochemical parameters. No significant adverse effects on renal function were observed across the tested doses (IC50, IC50 × 2, and IC50 × 3). While AST and ALT levels remained stable, ALP levels showed a dose-dependent modulation. These findings highlight the potential of 3g as a promising anticancer agent for glioblastoma, warranting further investigation into its detailed mechanisms of action and long-term safety.