DESIGN, SYNTHESIS, AND COMPREHENSIVE CHARACTERIZATION OF NOVEL PEMETREXED ANALOGUES FOR TARGETED NON-SMALL CELL LUNG CANCER THERAPY: A PRECISION-DRIVEN APPROACH TO ENHANCING CHEMOTHERAPEUTIC EFFICACY

Abstract

The synthesis and characterization of Pemetrexed analogues (PMA-1 and PMA-2) were performed to enhance the stability, permeability, and anticancer potential of the parent drug. The synthesized analogues exhibited higher melting points (PMA-1: 229–234°C, PMA-2: 235–239°C) compared to Pemetrexed (225–227°C), indicating increased thermal stability. PMA-1 had a higher yield (82%) than PMA-2 (74%), suggesting more efficient synthetic pathways. Thin Layer Chromatography (TLC) analysis revealed increased Rf values (PMA-1: 0.56, PMA-2: 0.59), suggesting improved lipophilicity. FTIR spectral analysis confirmed successful structural modifications, with shifts in C=O stretching (PMA-1: 1655–1685 cm⁻¹, PMA-2: 1660–1690 cm⁻¹) and N-H stretching (PMA-1: 3100–3500 cm⁻¹, PMA-2: 3200–3450 cm⁻¹), indicating altered hydrogen bonding and electronic interactions. Mass spectrometry analysis further confirmed the successful synthesis, with M+ ion peaks at m/z 563.54 for PMA-1 and m/z 577.56 for PMA-2. Sulforhodamine B (SRB) assay was used to test the analogues' anticancer potential against the following cell lines: HeLa (cervical cancer), A549 (lung cancer), MCF-7 (breast cancer), and HCC1937 (breast cancer). PMA-1 consistently exhibited stronger cytotoxic activity than PMA-2, demonstrating dose-dependent inhibition across all cell lines. At 90 μg/mL, PMA-1 achieved 84.55% inhibition in HCC1937, 87.22% in MCF-7, 82.15% in A549, and 81.79% in HeLa cells, outperforming PMA-2. These findings suggest that PMA-1 may be a superior candidate for further anticancer evaluation, with potential improvements in drug stability, solubility, and therapeutic efficacy. Further in vivo studies and mechanistic investigations are warranted to validate its clinical relevance.