Phytochemical Profiling and GC-MS Analysis of Annona muricata Ethanol Leaf Extract as potential agent for Nephrolithiasis management: An In-vitro and In-silico Investigation

Medicinal plants have long been utilized for therapeutic purposes; however, the specific effects and safety profile of ethanol Annona muricata leaf extract in nephrolithiasis management remain underexplored. This study aimed to evaluate the phytochemical composition, antioxidant activity, toxicity, and potential molecular mechanisms of the extract in relation to kidney stone formation. Phytochemical profiling was conducted using Gas Chromatography-Mass Spectrometry (GC-MS) and quantitative analysis of active constituents. Antioxidant potential was determined through in-vitro assays, including diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging and nitric oxide inhibition, to evaluate its free radical neutralizing capacity. Acute toxicity assessment provided insight into the extract's safety profile and potential therapeutic window. Furthermore, insilico investigations, comprising molecular docking and gene ontology analysis, were employed to elucidate the extract’s bioactive interactions at the molecular level, identifying potential mechanisms responsible for its nephroprotective effects. Bioactive compounds identified through GC-MS analysis were found to interact with molecular pathways and gene expressions linked to prevention and inhibition of kidney stone formation. A. muricata ethanol extract phytochemicals exhibited the strong binding to TNF-α and IL-1β with overall binding energy ranges from -3.6 to -6.3 kcal/mol for TNF-α and -4.1 to -5.8 kcal/mol for IL-1β. Findings from this study provide compelling evidence supporting the therapeutic potential of A. muricata in nephrolithiasis prevention and management, highlighting its promise as a natural remedy in kidney stone treatment.