Integrated experimental and computational insights into the anti-inflammatory potential of flower-derived exosome-like nanoparticles targeting the NF-κB pathway

Introduction: Dysregulated inflammation underlies numerous chronic pathologies, with the NF-κB p65-p50 heterodimer acting as a pivotal transcriptional regulator that mediates different inflammatory responses. Consequently, inhibiting NF-κB nuclear translocation has emerged as a promising strategy in anti-inflammatory drug development. While floral extracts have been widely used, recent advances have highlighted the therapeutic potential of flower-derived exosome like nanoparticles as promising cell-free therapeutics owing to their enhanced biocompatibility and stability. Methods: Exosome like nanoparticles were isolated from three ethnomedicinal flowers and systematically characterized. Antioxidant potential of ELNs was evaluated through DPPH assay and their anti-inflammatory potential was assessed. Further, to elucidate the molecular mechanisms underlying NF-κB modulation, key ELN-associated metabolites were computationally screened against the crystallized NF-κB p65-p50 heterodimer using molecular docking, followed by molecular dynamics simulations to evaluate binding stability and interaction dynamics. Results and discussion: analysis revealed significant regulation in mRNA expression of inflammatory cytokines and NF-κB transcriptional activity. Molecular docking identified several metabolites with higher binding affinity against NF-κB p65-p50 heterodimer supported by simulation studies confirming stable ligand-protein interaction. Both docking scores and simulation trajectories strongly supported stable, high-affinity interactions consistent with NF-κB pathway inhibition. Overall, the combined experimental and computational findings in the study represent the first comprehensive data of floral ELN bioactives, offering the significant translational potential of floral nanovesicles as a new class of biocompatible, cell-free nanotherapeutics for anti-inflammatory drug discovery.