LC-HRMS-Based Phytochemical Profiling of Azadirachta indica A. Juss Leaf Extract and In Silico Screening for Plasmodium vivax Dihydrofolate reductase (DHFR) Inhibitors
Year: 2026
Type: article
Abstract: Malaria persists as a major challenge to global health, and rising resistance of Plasmodium vivax to antifolate drugs necessitates the discovery of new antimalarial agents. This research investigated methanolic Azadirachta indica (neem) leaf extract by employing an integrated workflow combining liquid chromatography–high-resolution mass spectrometry (LC-HRMS) metabolite profiling, ADMET prediction, and molecular docking (MD) against Plasmodium vivax dihydrofolate reductase (PvDHFR). A methanolic leaf extract was prepared and subjected to qualitative phytochemical screening, determination of total phenolic, flavonoid, and condensed tannin contents, and LC-HRMS metabolite profiling. Annotated metabolites were evaluated for physicochemical, pharmacokinetic, and toxicity properties, followed by MD against wild-type PvDHFR (PDB ID: 2BL9). The extract contained flavonoids, phenolics, alkaloids, steroids, triterpenoids, and terpenoids. Total phenolic and flavonoid contents were 40.09 ± 0.10 mg GAE/g and 11.22 ± 0.35 mg QE/g, respectively, while condensed tannins were below the detection limit. LC-HRMS tentatively identified 19 metabolites, with quercetin (22.13%) and quercetin-3β-D-glucoside (14.32%) as the dominant constituents. Docking validation showed excellent reliability with an RMSD of 0.295 Å. Among the annotated compounds, pinostrobin showed the strongest predicted binding affinity for PvDHFR (-7.7 kcal/mol), slightly better than pyrimethamine (-7.6 kcal/mol), whereas pinocembrin and alpinetin also demonstrated favorable docking scores (-7.6 kcal/mol). Aglycone flavonoids displayed superior drug-likeness, pharmacokinetic, toxicity, and docking profiles compared with glycosylated flavonoids and triterpenoids. These findings suggest that LC-HRMS-guided computational screening can effectively prioritize neem-derived compounds for antimalarial drug discovery and provide a rational foundation for subsequent enzyme inhibition assays, in vitro antiplasmodial evaluation, and safety assessment in future studies.
Institution: Universitas Islam Malang
Language: English
Subfield: Drug Discovery
Domain: Life Sciences
Open Access status: diamond
OpenAlex ID: W7206202829