Biological and In Silico Studies of a Novel Pyrazolo[3,4-d]Thiazole Derivatives: Anticancer, Anti-Inflammatory,...

This study synthesized and evaluated a series of novel 1,3,5-trisubstituted-1H-pyrazolo[3,4-d]thiazole derivatives (5a, 5b, 6a, 6b, and 7) for their cytotoxic, anti-inflammatory, and anticancer properties. The cytotoxic activity of these derivatives was evaluated against MCF-7 and HepG2 cell lines. Compound 6b demonstrated the most potent anticancer activity, with IC₅₀ values of 15.57 ± 2.93 µg/mL for MCF-7 cells and 43.72 ± 1.90 µg/mL for HepG2 cells, demonstrating greater efficacy than doxorubicin. Mechanistic studies revealed that compound 6b induced S-phase arrest in MCF-7 cells and G1/S-phase arrest in HepG2 cells, along with a significant increase in apoptosis rates in the treated cancer cells. Moreover, compound 6b demonstrated significant vascular endothelial growth factor receptor 2 (VEGFR-2) inhibition, surpassing the efficacy of sorafenib. Compounds 5b and 6b demonstrated significant anti-inflammatory activity in RAW264.7 macrophage cells, as indicated by the decrease in nitric oxide (NO) production and the downregulation of proinflammatory cytokines, such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). The derivatives exhibited no cytotoxicity toward normal cell lines (MCF-10A, THLE-2, and Vero cells), as validated by MTT assays. Docking studies elucidated the interaction mechanism between the compounds and the enzyme receptors. These findings demonstrate compound 6b as a promising candidate for the development of dual-function anticancer and anti-inflammatory agents.