Exploring the HNE-inhibitory potential of in silico selected Moringa oleifera defense proteins.

Human neutrophil elastase (HNE) is a primary therapeutic target for the treatment of severe inflammatory disorders. This study explores the HNE-inhibitory potential of peptides derived from Moringa oleifera 2S albumin and chitin-binding proteins, leveraging the inherent proteolytic stability of these defense-related scaffolds. Using a pipeline-assisted approach, mass spectrometry-based proteomics was combined with in silico proteolysis and molecular docking to guide the investigation. Enzyme kinetic studies established a competitive mode of inhibition, confirming the predicted binding mechanism. Initial virtual screenings utilized porcine pancreatic elastase as a structural surrogate, followed by rigorous docking simulations against the HNE catalytic site to evaluate clinical applicability. Molecular docking identified key residues at the protein-ligand interface that are likely responsible for stabilizing the complex across multiple subsites of the HNE pocket. Assays with chemically synthesized lead peptides corroborated the computational predictions, demonstrating notable HNE-inhibitory activity. These findings present an integrated computational-experimental workflow for the rapid identification of plant-derived inhibitors and offer promising, stable lead scaffolds for the development of scalable, next-generation anti-inflammatory agents.