Design, synthesis and biological evaluation of testosterone derivatives as potential anti-tumor and anti-inflammatory agents.

A series of 29 novel testosterone derivatives (14a-14k and 15a-15r) containing substituted 1,2,3-triazole groups were designed and synthesized via click chemistry. Their anticancer and anti-inflammatory activities were evaluated in vitro and in vivo. Among these, compound 15b showed the most potent antiproliferative effect against HepG2 (IC₅₀ = 4.84 μM), while exhibiting low toxicity toward normal L02 cells (IC₅₀ > 40 μM). Mechanistic studies revealed that 15b induced DNA damage via upregulation of γ-H2AX, disrupted cell cycle progression, and triggered mitochondrial-mediated apoptosis through downregulation of caspase-9, caspase-3, and PARP. Molecular docking suggested a possible interaction with human topoisomerase II, indicating that it could be a potential target contributing to the DNA-damaging activity. In vivo, 15b significantly inhibited HepG2 xenograft growth without affecting organ indices. In addition, 15b displayed strong anti-inflammatory activity in LPS-stimulated BV-2 microglia, reducing mRNA and protein levels of IL-1β, IL-6, TNF-α, iNOS, COX-2, and NLRP3, indicating multi-target inhibition of neuroinflammation. In summary, compound 15b exhibits dual anticancer and anti-inflammatory activities by inducing DNA damage, activating apoptotic pathways in tumor cells, and suppressing key pro-inflammatory mediators, highlighting its potential as a lead compound for the development of anticancer drugs with anti-depressive properties.