Identifying Anti‐NSCLC Bioactive Compounds in Scutellaria via 2D NMR‐Based Metabolomic Analysis of Pharmacologically Classified Crude Extracts

Abstract We presented a strategy utilizing 2D NMR‐based metabolomic analysis of crude extracts, categorized by different pharmacological activities, to rapidly identify the primary bioactive components of TCM. It was applied to identify the potential bioactive components from Scutellaria crude extracts that exhibit anti‐non‐small cell lung cancer (anti‐NSCLC) activity. Four Scutellaria species were chosen as the study subjects because of their close phylogenetic relationship, but their crude extracts exhibit significantly different anti‐NSCLC activity. Cell proliferation assay was used to assess the anti‐NSCLC activity of four species of Scutellaria . 1 H− 13 C HSQC spectra were acquired for the chemical profiling of these crude extracts. Based on the pharmacological classification (PCA, OPLS‐DA and univariate hypothesis test) were performed to identify the bioactive constituents in Scutellaria associated with the anti‐NSCLC activity. As a result, three compounds, baicalein, wogonin and scutellarin were identified as bioactive compounds. The anti‐NSCLC activity of the three potential active compounds were further confirmed via cell proliferation assay. The mechanism of the anti‐NSCLC activity by these active constituents was further explored via flow cytometry and western blot analyses. This study demonstrated 2D NMR‐based metabolomic analysis of pharmacologically classified crude extracts to be an efficient approach to the identification of active components of herbal medicine.