Cilianthaxone, a novel compound isolated from ethanolic extract of Ephedra ciliata and exploring its anti-inflammatory potential through in vitro, in vivo, and in silico studies.

The current study was designed to determine the anti-inflammatory potential of a novel compound isolated from ethanolic extract of Ephedra ciliata (ECE). The extract was fractionated and purified using various chromatographic techniques, followed by structural elucidation via spectroscopic methods. ECE and isolated compound cilianthaxone (ECE-2e-4) were analysed for acute toxicity by in vivo study for 14days and it was found that a dose of 2000mg/kg has no significant changes on animal body normal parameters. In vivo, animal studies like a hot plate and acetic acid-induced writhing methods were applied to determine the analgesic potential of both the ECE and cilianthaxone. In the hot plate, carrageenan-induced paw edema model, the cilianthaxone exhibited greater anti-inflammatory potential at a dose of up to 10mg/kg compared to the standard drug indomethacin. Cilianthaxone exhibited 85.35% inhibition compared to indomethacin, which showed 77.61% inhibition. The analgesic activity of the isolated compound was also confirmed by the acetic acid-indsuced writhing method, where cilianthaxone exhibited a delayed latency time of 30.50min, comparable to the standard drug morphine. Antioxidant properties of cilianthaxone were also confirmed through DPPH and NO scavenging methods and have IC50 values of 14.28±0.43μg/kg and 15.60±0.21μg/kg, respectively. The chorioallantoic membrane (CAM) assay was also performed to check anti-angiogenesis, and no angio-toxicity was found. Furthermore, in silico studies were also performed to determine the binding mode and protein targeting ability of isolated compound. Overall, the current research highlights the significant anti-inflammatory effects and wound-healing potential of cilianthaxone, supported by its favorable target protein interactions.