Targeting microtubule dynamics: synthesis and evaluation of novel 4-thiazolidinone derivatives in pancreatic Cancer

Pancreatic Cancer remains one of the most lethal malignancies, with limited therapeutic options and high chemoresistance partly associated with β-tubulin overexpression. In this study, we synthesized and biologically evaluated a series of 4-thiazolidinone derivatives, identifying Les-6666 and Les-5549 as potent Anticancer candidates. An optimized microwave-assisted synthetic protocol enabled efficient and environmentally friendly production of selected derivatives. In silico ADMET profiling predicted favorable drug-like properties, high gastrointestinal absorption, and no blood-brain barrier penetration. Molecular docking studies revealed strong binding affinity of selected compounds to the colchicine and combretastatin A4 (CA4) sites of β-tubulin. Biological evaluation through the NCI-60 screening program demonstrated pronounced antiproliferative activity, with Les-6666 exhibiting submicromolar GI₅₀ values across multiple Cancer cell lines. In pancreatic MIA PaCa-2 cells, both Les-6666 and Les-5549 showed potent cytotoxicity (IC₅₀ = 0.08 μM and 0.10 μM, respectively), inhibited DNA synthesis, suppressed clonogenic survival, reduced migration, arrested cells at the G₂/M phase of the cell cycle and induced Apoptosis. Tubulin polymerization assays confirmed significant depolymerizing activity comparable to CA4. Furthermore, Les-6666 reduced β-tubulin protein expression, consistent with tubulin autoregulatory feedback mechanisms. Collectively, these findings identify Les-6666 as a promising antiproliferative agent that inhibits tubulin polymerization in pancreatic Cancer cells warranting further investigation.