Synthesis, characterization, biological evaluation, and molecular modeling of novel nimesulide urea derivatives as potential MetAP2 inhibitors

A series of nimesulide-derived ureas were synthesized in high yields (69- 91%) via a novel synthetic method under organotin catalysis. The structural characterization of the synthesized compounds was determined using a variety of spectroscopic methods, including ¹H NMR, ¹³C NMR, FTIR, and HRMS. Following initial screening, selected compounds were further evaluated using specific biological assays, including cytotoxicity analysis, apoptosis assessment by Annexin V/PI staining, analysis of Bax and Bcl-2 protein expression, and detection of DNA fragmentation using AO/EB staining. These evaluations were carried out in a variety of cell lines, including MDA-MB-231 (human triple-negative breast cancer cells), HeLa (human cervical cancer cells), PC-3 (androgen-independent human prostate cancer cells), MKN-45 (human gastric cancer cells), U87 (human glioblastoma cancer cells), and HUVEC (human umbilical vein endothelial cells). Three of the compounds demonstrated efficacy in MDA-MB-231 cells, resulting in increased AO/EB staining and annexin-V-PI binding levels, and increased Bax/Bcl-2 ratios. Numerous studies implicate MetAP2 in angiogenesis. MetAP2 stimulates cancer cell proliferation when it is upregulated and appears to play an essential role in tumor progression. In connection with the cell lines studied in this study, a crucial MetAP2 enzyme target was selected for in silico studies to support the experimental outcomes. The three promising compounds have been demonstrated to accelerate cell apoptosis and inhibit cell division by targeting MetAP2. The compounds synthesized in this study potential to overcome the challenges of targeted therapies in triple-negative breast cancer. Alongside these findings, the antioxidant potential of the synthesized compounds was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. One of the compounds exhibited a radical scavenging activity comparable to the reference standard. In-silico molecular modeling studies were conducted in the final phase to evaluate the newly designed inhibitors as potential anticancer and antioxidant drug candidates. The most likely conformations of the MetAP2-ligand complex were sorted according to the free energy of binding score (kcal/mol) and agree with the experimental inhibition values. The computational results suggest that the newly synthesized compounds may be potential anticancer drug candidates.