Phytochemical profiling and pharmacological evaluation studies of anxiolytic efficacy of <i>Glycyrrhiza glabra</i> root decoction

Glycyrrhiza glabra ( G. glabra) root is traditionally known for its wide-ranging pharmacological properties, including antimicrobial, anti-inflammatory, hepatoprotective, and immunomodulatory effects. This study aims to explore the anxiolytic potential of G. glabra root decoction. Using LC-MS, seven major phytocompounds such as N-acetylalanine, primuliten, 8-hydroxy-13,14,15,16-tetranor-12-labdanoic acid, gardnerine, abrine canrenoic acid, and aspterric acid were identified from the G. glabra decoction. These compounds were further evaluated through ADME profiling and network pharmacology to assess their drug-likeness and pharmacokinetic properties using the SWISS ADME platform. Among the identified compounds, the bioactive compound canrenoic acid was found to be investigated for the direct modulation of ion channels in the brain contributing to central nervous system (CNS) pharmacology, and for the regulation of CNS stress responses, which can modulate anxiety-related behaviours. Canrenoic acid (PubChem CID: 656615) demonstrated promising interactions with the anxiety-related protein target PDB ID:7E2Y. Molecular dynamics (MD) simulations were conducted over a 100-nanosecond trajectory to analyze the stability of the Canrenoic acid–7E2Y complex. The simulation results revealed minimal conformational fluctuations, with the ligand maintaining a stable binding pose through hydrophobic interactions and hydrogen bonding. root mean square deviation (RMSD), root mean square fluctuation (RMSF), secondary structure evolution, and protein-ligand interaction analyses confirmed the robustness of the complex. In addition, in vivo testing using the elevated plus maze (EPM) was performed in rats to evaluate the anxiolytic effects of four different extracts at two dose levels (100 mg/kg and 200 mg/kg), compared to a standard treatment group (diazepam 2 mg/kg). The G. glabra decoction exhibited significant anxiolytic activity. Rats treated with 200 mg/kg of G. glabra showed a marked increase in both the number of entries into the open arms (3.33 ± 0.21) and the time spent in the open arms (43.83 ± 0.79 seconds), closely mirroring the effects observed with the standard drug diazepam. These findings suggest that canrenoic acid is a promising candidate for further preclinical and clinical evaluation in the treatment of anxiety disorders.