RP-HPLC quantification and separation of isomeric bacosides and exploration of their anti-schizophrenic potential by...

Schizophrenia is a chronic and heterogeneous neuropsychiatric disorder involving positive, negative, and cognitive symptoms. Antipsychotic drugs prescribed currently are effective in controlling positive symptoms but offer limited benefits for negative and cognitive impairments as they focus on single-target or narrow-mechanism treatment strategies without sufficiently addressing the multi-factorial dysfunction underlying schizophrenia. Therefore, multi-target natural compounds like Bacopa monnieri (Brahmi) are increasingly being explored as potential therapeutic candidates for neurological disorders like schizophrenia. However, despite its reported neuroprotective and cognition-supporting properties, the clinical translation of Brahmi remains limited due to the variability in bacoside content across formulations and insufficient understanding of their potential molecular interactions in schizophrenia-associated pathways. In the present study, a reliable HPLC chromatographic method was developed and validated for the simultaneous quantification of major isomeric bacosides, i.e., bacopaside I, bacopaside II, bacoside A3, bacopaside X, and bacopasaponin C, in multiple batches of different formulations. The developed method demonstrated acceptable linearity, accuracy, and sensitivity, enabling consistent bacoside quantification and revealing formulation and batch-dependent variations. To further explore the potential pharmacological relevance and understand the molecular interactions of these bacosides, network pharmacological analysis was performed. Our analysis revealed 97 common targets for the five bacosides, of which 76 were schizophrenia-specific targets. The serotonergic pathway emerged as a key candidate with 5-HT1A, the serotonin receptor, as the top target for bacosides. Molecular docking studies with 5-HT1A confirmed favourable binding poses of bacosides, and bacopasaponin C had the highest docking score. The 50 ns molecular dynamics simulation of the 5HT1A-bacopasaponin C complex demonstrated stable protein-ligand interactions throughout the simulation period. Overall, this study provides an integrated framework combining analytical chemistry with exploratory computational analysis for the anti-schizophrenic potential of Brahmi.