In silico and in vitro evaluation of small oligopeptide derived from CfTX-A toxin for its anti-inflammatory activity against LPS/MSU-induced fibroblast cells

Abstract

Venoms contain a wide range of biologically active peptides with multiple pharmacological properties, making them important for novel therapeutic development. This study involves in silico and in vitro approaches to evaluate the anti-inflammatory potential of a small oligopeptide derived from jellyfish Chironex fleckeri toxin CfTX-A. Using computational methods, structural modeling, and ADMET screening, the active peptides were identified based on their physicochemical properties, enzymatic stability, and predicted interaction with key inflammation-associated proteins such as NLRP3, caspase-1, xanthine oxidase (XO), and IL-1β. Molecular docking predicted high-affinity binding of peptide WPNY to active sites within these targets, suggesting its potential dual role in modulating inflammatory and uric acid pathways. The peptide further demonstrated potent XO inhibition (47.98 ± 12.41%) which was comparable to allopurinol (Alp) drug control. In LPS/MSU-stimulated fibroblast cells, CfTX-A peptide at 125 µM maintained high cytocompatibility (97.23 ± 3.83%), significantly suppressed NLRP3 inflammasome activation (1.23 ± 0.07), NO (15.35 ± 1.77 μM) and IL-1β (26.86 ± 3.01 pg/ml) production, NF-κB phosphorylation (1.52 ± 0.06), and downstream inflammatory mediators including iNOS (2.88 ± 0.10), COX-2 (6.24 ± 0.67), TNF-α (2.85 ± 0.16), and MMP-9 (4.06 ± 0.38). It also reduced intracellular ROS levels, retained thermal stability, and showed efficient uptake within the cells. These findings suggest the peptide’s dual role as an anti-inflammatory and antioxidant agent. Further, it demonstrates the utilization of venom-derived peptides and in silico-driven platforms in accelerating peptide drug discovery. Future in vivo investigations in arthritis models are essential to establish therapeutic relevance.