Exploration of 1-(2,4-difluorophenyl)-5-oxopyrrolidine-3-carboxylic acid derivatives effect on triple-negative breast, prostate cancer and melanoma cell 2D and 3D cultures

Abstract

1-Substituted 5-oxopyrrolidine-3-carboxylic acid and its derivatives play an important role as components of many biologically active molecules. This study describes the synthesis of 1-(2,4-difluorophenyl)-5-oxopyrrolidine-3-carboxylic acid derivatives and their anticancer properties. The target compounds were prepared using 2,4-difluoroniline as a starting material; in this way, derivatives of benzimidazoles, hydrazones and azoles were formed. Investigation of the anticancer activity of all synthesized compounds showed that the hydrazones had the strongest effect on cancer cell lines. Compounds were tested for their cytotoxic effect by the MTT assay in human triple-negative breast cancer MDA-MB-231, prostate adenocarcinoma PPC1, melanoma A375 and human foreskin fibroblasts CRL-4001 after 72 hours of incubation. The impact of the compounds on cancer cell migration was assessed using a 'wound healing assay'. Activity in 3D cultures was determined by evaluating changes in spheroid size and assessing cell viability. Overall, the selected compounds 7b, 9c, 9e, 9f and 10 exhibited greater activity in the A375 cell line and were less active against the MDA-MB-231 cell line. Compounds 9c, 9e and 10 showed relatively higher selectivity for cancer cells over fibroblasts. Hydrazone 9f, bearing N'-(4-methylbenzylidene) moiety, was identifiedasthe most cytotoxic compound in both prostate adenocarcinoma PPC-1 and melanoma A375 cells in monolayer and 3D culture models. Compound 9e, with N'-(4-bromobenzylidene) moiety, exhibited the most pronounced inhibitory effect on cell migration as determined by the 'wound healing' assay.