Elucidating the Multi-Target Mechanisms of Niclosamide in Hepatocellular Carcinoma through Network Pharmacology, Molecular Docking, and Molecular Dynamics Simulations

The Hepatocellular carcinoma (HCC) continues to be a major contributor to cancer-related deaths globally, fueled by intricate molecular diversity and widespread resistance to existing targeted treatments. Drug repurposing provides a strategy that is both effective and economical for identifying new therapeutic options. Niclosamide (NIC), which is approved by the food drug administration for use as an anthelmintic, has shown promise as a cancer treatment because it can affect multiple oncogenic signalling pathways. This study utilized an integrated in silico approach that combined network pharmacology, molecular docking, molecular dynamics (MD) simulations, and toxicity prediction to clarify the multitarget mechanisms of NIC in HCC. The analysis using network pharmacology found 252 overlapping targets between genes associated with NIC and those linked to HCC. The examination of protein–protein interactions underscored essential hub proteins such as AKT1 (Protein Kinase B alpha), Epidermal Growth Factor Receptor (EGFR), Proto-oncogene Tyrosine-Protein Kinase Src (SRC), Signal Transducer and Activator of Transcription 3 (STAT3), Janus Kinase 2 (JAK2), and Phosphoinositide-3-Kinase Regulatory Subunit 1 (PIK3R1). This indicates that NIC may play a role in vital oncogenic signalling networks. Analyses of Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment showed a significant association with pathways such as Phosphoinositide 3-Kinase (PI3K)–AKT, ERBB, JAK–STAT, EGFR inhibitor resistance, and immune-related pathways. This highlights the polypharmacological nature of NIC. Molecular docking showed that NIC exhibits favorable binding affinities toward major hub targets, similar to those of the clinically used drug Sorafenib (SOR). Additionally, 100-ns MD simulations verified the dynamic stability and ongoing interactions of NIC with essential proteins like AKT, EGFR, and SRC, suggesting strong binding under physiological conditions. In silico toxicity assessment via the ProTox-III server indicated low to moderate toxicity levels, with hepatotoxicity considered acceptable and only minimal risks for mutagenicity or carcinogenicity. This extensive computational investigation endorses niclosamide as a multitarget, mechanistically sound, and relatively safe candidate for HCC therapy, thereby necessitating further experimental validation and formulation optimization.