Deciphering the immunomodulatory mechanisms of piperine during HIV-Mtb co-infection: network pharmacology, docking and molecular dynamics.

UNLABELLED: Human immunodeficiency virus (HIV) and Mycobacterium tuberculosis (Mtb) co-infection synergistically weakens the host immune system. Previous studies have investigated the involvement of multiple signalling pathways and genes such as TNF, IL6, PI3K-AKT1, and JAK/STAT3 in regulating host immune functions during HIV and Mtb infection. These pathways have also been explored for apoptosis, autophagy, and anti-neoplastic activity of several bioactive compounds, including piperine, the major alkaloid of Piper nigrum. However, the therapeutic potential of piperine in modulating host immune responses during HIV/Mtb co-infection has not been studied. Therefore, the present investigation aims to elucidate the potential immunomodulatory role of piperine during HIV/Mtb co-infection using network pharmacology, docking and molecular dynamics (MD) simulations. The gene targets related to HIV, Mtb, and piperine were retrieved from public databases. Common gene targets were identified using Venny 2.0.2/Appyters. Protein-protein interaction (PPI) networks were constructed via STRING and analysed with Cytoscape to determine hub genes. Gene Ontology (GO) enrichment and KEGG pathway analyses were carried out using SRplot. Our study identified 78 potential therapeutic targets and 20 hub genes associated with piperine. Among these, TNF, IL6, AKT1, STAT3 and CASP3 emerged as central targets across the top signalling pathways during HIV/Mtb co-infection. Molecular docking demonstrated strong binding affinities of piperine with host, HIV, and Mtb targets, showing binding energies between - 6.0 and - 9.2 kcal/mol. Furthermore, 100 ns molecular dynamics simulations confirmed the structural stability of the selected protein-ligand complexes, exhibiting low structural deviations, limited residue fluctuations, preserved protein compactness, and persistent hydrogen-bond interactions throughout the simulation period. These findings suggest that piperine could serve as a promising immunomodulatory compound in the treatment of HIV/Mtb co-infection. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at https://doi.org/10.1007/s40203-026-00754-w.