Liposomal formulation of Fagonia arabica L. enhances antithrombotic efficacy: phytochemical, pharmacological, and computational investigations

Abstract

Fagonia arabica “Dhamasa” is traditionally used in Ayurvedic and Unani medicine across South Asia and the Middle East for its therapeutic properties, notably as a natural blood purifier. It is believed to aid in dissolving blood clots and reducing the risk of brain hemorrhage and cardiovascular events. Given previous reports of saponin content in F. arabica, this study aimed to evaluate its thrombolytic and anticoagulant potential. After confirming its saponin richness through NMR (1D, 2D) and LC-MS, we developed liposomal nanoparticles to enhance bioavailability and target clot dissolution more effectively. Liposomes were characterized using TEM, particle size, PDI, and zeta potential. In vitro thrombolytic and anticoagulant activities were assessed, followed by in vivo testing in adult Wistar rats. The dose-dependent effects of the saponin-rich n-butanol fraction of F. arabica on Antithrombin-III levels were evaluated using an in vitro quantitative immunoturbidimetric assay. Coagulation parameters were evaluated using aPTT and PT assays. A molecular docking study was systematically performed on nine targets using the five structurally elucidated steroidal and terpenoid glycosides to predict and evaluate their binding affinities. These computational findings were designed to complement ongoing in vitro and in vivo investigations. The butanol extract showed significant in vitro thrombolytic and anticoagulant effects at 40 mg mL⁻¹. The results demonstrate that the saponin-rich fraction modulates Antithrombin-III in a concentration-dependent manner. Liposomal nanoparticles achieved similar efficacy in vivo at just 3 mg kg⁻¹, compared to 100 mg kg⁻¹ for the non-nano extract. The liposomal group showed PT of 35 ± 4 s and aPTT >180 s, outperforming the non-nano extract (PT 30 ± 7 s, aPTT >180 s) and controls (PT 12 ± 2 s, aPTT 37 ± 3 s). The docking study provides mechanistic insight into their potential multi-target antithrombotic activities. Besides the butanol extract, liposomal F. arabica enhances antithrombotic efficacy at lower doses, supporting its potential as a natural therapeutic candidate for thrombotic disorders.