New 3H-1,2-Dithiole-3-thione derivatives: Design and synthesis, H2S-release profile, in vitro anticancer activity, and in silico multi-target assessment

Abstract Cancer continues to be a leading cause of global mortality, highlighting the ongoing need for novel anticancer compounds that offer high efficacy with improved side effect profiles. In the present study, a series of 3 H -1,2-dithiole-3-thione derivatives ( DTT-S1-18 ) were synthesized as promising anticancer agents, and the structures of products were confirmed by spectral techniques. H 2 S-releasing experiments showed that most of the compounds released higher amounts of H 2 S slowly over time compared to standard ADT-OH . All compounds were tested for antiproliferative activity on HT-29, PC-3, MCF-7, and HUVEC cell lines. Compounds DTT-S6 (3-nitrophenyl derivative) and DTT-S8 (methionine derivative) have the lowest IC 50 values of 41.6 and 38.9 µM on the MCF-7 cell line, respectively. Based on the wound healing and colony formation assays performed in MCF-7 cells, the wound areas were not significantly changed after treatment with compounds DTT-S6 and DTT-S8 , whereas compound DTT-S8 at double IC 50 dose inhibited colony formation by 81.82%. In addition, molecular docking, MD simulations, MM/GBSA binding free energy calculations, and binary QSAR analyses were performed to explore the potential target interactions and predicted activity profiles of the synthesized compounds toward inflammation-related proteins, including COX-1, COX-2, 5-LOX, and iNOS, thereby supporting the development of mechanistic hypotheses for future validation. Furthermore, structure–activity relationship (SAR) analyses were conducted to correlate the structural characteristics of the synthesized compounds with their H 2 S releasing potential and biological profiles. Overall, this work integrates experimental anticancer evaluation with computational pathway and structure-based cancer/inflammation analyses to characterize novel DTT-based H 2 S donors. The findings identify particularly compound DTT-S8 , as a promising in vitro anticancer candidate, while the computational results suggest a putative involvement of inflammation-related targets, particularly the COX-2/5-LOX axis, which requires direct biochemical and cellular validation.