Anti-proliferative and anti-inflammatory potential of 2-chloro-3-((4-hydroxyphenyl)amino)naphthalene-1,4-dione through targeting TNF-α/IL-8 and NF-κB signaling in HCT116 cancer cells: a comprehensive in silico docking and in vitro study

Abstract

Colorectal cancer (CRC) is closely associated with chronic inflammation, and the NF-κB pathway acts as a central regulator of the inflammatory response that promotes tumor progression. In this study, we examined the anticancer and anti-inflammatory potential of anilino-1,4-naphthoquinone derivatives (compounds 3, 8, and 12) in CRC models using in vitro assays, in silico analysis with the Gold program, and molecular dynamics (MD) simulations. HCT116 cells were pre-exposed to these compounds before LPS stimulation. Cytotoxicity was determined using the MTT assay. The results indicated that compounds 3, 8, and 12 inhibited cell proliferation, leading to nuclear morphological changes, as evidenced by an increased sub-G1 population in HCT116 cells. In addition, Flow cytometry analysis showed that the compound significantly increased apoptotic cell death, particularly in the late-apoptotic population, compared with the control and reference treatments. Molecular docking analysis demonstrated that the tested compound favorably interacted with key amino acid residues within human NF-κB/p50, the p50/p65 NF-κB complex, and IKKβ, with binding patterns to the active sites comparable to those of the reference inhibitors, MLN120B and BMS-345541. In biological experiments, compound 12 reduced TNF-α and IL-8 secretion, indicating suppression of the NF-κB-mediated inflammatory pathway. Collectively, these findings suggest that compound 12 exerts anticancer activity through dual anti-inflammatory and pro-apoptotic mechanisms, possibly by modulating the NF-κB pathway. This compound may therefore represent a promising lead candidate for further drug development against CRC.