Untargeted metabolomics approach based on UPLC-ESI-QTOF-MS/MS and evaluation of antioxidant and cytotoxic activities of Nepeta deflersiana Schweinf. ex Hedge supported by molecular modeling and ADMET analysis

Abstract

Nepeta deflersiana is a traditional medicinal plant in Saudi Arabia, recognized for its therapeutic properties. This study aimed to identify the phytochemicals present in its crude extract and five fractions, and repectively, evaluate their antioxidant potential using DPPH and ABTS assays, and assess their cytotoxicity against human breast adenocarcinoma (MCF-7) and colorectal adenocarcinoma (HT-29, SW-480) cell lines. In addition, ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) was applied for metabolomic profiling of the most potent extract. The results showed significant total phenolic (30.58 ± 0.10 mg GAE/g) and flavonoid (26.30 ± 1.06 mg rutin equivalent/g) contents in the crude extract. The aqueous fraction exhibited the strongest antioxidant activity, with IC50 values of 0.256 ± 0.006 mg/mL (DPPH) and 1.225 ± 0.010 mg/mL (ABTS). It also demonstrated notable cytotoxic effects, particularly against MCF-7 (5.92 ± 0.29 μg/mL) and HT-29 (4.83 ± 0.87 μg/mL) cell lines. UPLC-QTOF-MS/MS analysis identified 35 compounds, including phenolics, flavonoids, amino acids, and peptides. Subsequently, molecular docking analysis of the major identified bioactive compounds with anticancer targets confirmed strong binding affinities. ADMET assessement of key bioactive compounds revaeled favorable pharmacological profiles and good drug-likeness properties. These findings highlight N. deflersiana as a promising natural source for anticancer drug development.

Molecular docking studies supported the cytotoxicity findings