Unveiling the immunomodulatory mechanisms of pineapple metabolites: A multi-modal computational analysis using network pharmacology, molecular docking, and molecular dynamics simulation

The exploration of the immunomodulatory potential of pineapple metabolites holds promise for substantial implications across several fields, encompassing medicine, pharmacology, nutrition, and public health. This study explores potential immune-regulating properties of secondary pineapple metabolites beyond bromelain, using computational techniques. Pineapple juice's secondary metabolites were identified via LC-MS-based metabolomics, selected using KNApSAcK Kanaya and Dr. Duke's databases. A heatmap was generated with Orange v.3.27.0. Bioactivity predictions utilized the PASS Online webserver, while ADMET properties were forecasted. Network pharmacology explored metabolite interactions with protein targets. Molecular docking focused on compounds against receptors, choosing robust interactors for dynamic simulations. Our findings revealed that certain metabolites from pineapple juice/fermented juice exhibited interactions with proteins associated with pro-inflammatory cytokines. Specifically, the molecular docking results indicated that the carbohydrate moiety of bromelain (CMB) interacted strongly with TLR2, while 9,10-Dihydroxystearic acid showed strong interactions with NLRP3 (inflammasome). The flexibility and stability of these complexes were further assessed through molecular dynamics simulations conducted over a 50-ns time period. The MM-PBSA calculations also indicated low binding free energies between these complexes, suggesting strong molecular interactions. These findings suggest that CMB may interact with TLR2 and 9,10-Dihydroxystearic acid may interact with NLRP3, highlighting their potential as immunomodulatory agents. However, further experimental studies are warranted to confirm the therapeutic efficacy of these molecules and investigate their mechanisms of action in vivo.