Integrated In Silico and In Vivo evaluation of Cassytha filiformis: Molecular insights into caspase-3/8 binding and wound healing effect

Cassytha filiformis is a parasitic vine traditionally used in Chinese traditional medicine (CTM) for its anti-inflammatory, detoxifying, and wound-healing properties. Despite its ethnomedical significance, scientific validation of its therapeutic mechanisms-particularly its interaction with apoptotic pathways-remains limited. This study investigated the wound healing potential of Cassytha filiformis through in vivo and in silico approaches, alongside phytochemical isolation and characterization. Successive solvent extraction was performed using petroleum ether, chloroform, ethyl acetate, and methanol. Phytochemical screening confirmed the presence of flavonoids, phenolics, and phytosterols. Isolation and spectroscopic characterization identified pseudohypericin as key constituents. Gas chromatography-mass spectroscopy (GC-MS) analysis revealed additional 25 bioactive compounds. Among all extracts, the 5% chloroform extract showed the most significant activity in excision and incision wound models, achieving 100% wound contraction by day 16 and a tensile strength of 425.5 ± 4.89 g, outperforming the standard drug framycetin. In silico docking result showed strong binding affinity of quercetin 3-O-rutinoside (−11.9 kcal/mol) and pseudohypericin (−10.0 kcal/mol) with Caspase-3, supported by favorable ADMET and toxicity profiles. Similarly, Quercetin 3-O-rutinoside and Pseudohypericin exhibited significant binding affinities against Caspase-8 with binding score of -8.1 and -8.0 kcal/mol respectively. Both phytoconstituents showed significant binding affinity against Caspase-8. The stability of complex was validated through 100 ns molecular dynamic (MD) simulation results. Cassytha filiformis demonstrated potent wound healing activity, suggesting its potential for development into safe, effective plant-based therapeutics.