Metabolomic Fingerprinting of Cissampelos Sympodialis Eichler Leaf Extract and Correlation with Its Spasmolytic Activity.

Ethnopharmacological relevance: Cissampelos sympodialis Eichler (Menispermaceae) is popularly used in north-eastern Brazil for the treatment of respiratory diseases such as bronchitis and asthma. Despite many pre-clinical pharmacological studies, the compounds mediating the anti-asthma activity of polar extracts of Cissampelos sympodialis leaves have not been definitively identified. Aim of the study: Aim of the study: The aim of the study was to investigate the correlation between the bioactivity of polar extracts prepared from the leaves of C. sympodialis and the chemical composition using a H-1-NMR-based metabolomics approach. Material and methods: The metabolic profile of the leaf polar extract during different phenological stages of the plant was investigated using H-1 NMR spectroscopy while simultaneously screening for spasmolytic activity using guinea-pig tracheal preparations. The content of the alkaloids previously implicated in the bioactivity of Cissampelos sympodialis was determined by HPLC. Results: PCA analysis of the H-1 NMR data discriminated the extracts from different plant phenological stages. The contents of the major alkaloids decrease (from 2 +/- 0.32 mu g/mL for warifteine and 1 +/- 0.14 mu g/mL for methylwarifteine) to undetectable levels from 90 (CsL90 extract) and 120 (CsL120) days onwards for warifteine and methylwarifteine, respectively. All six extracts relaxed the trachea pre-contracted with carbachol, but the CsF210 extract was more potent (EC50 = 74.6 +/- 7.9 mu g/mL) compared to both CsL90 extracts and CsL i80 in the presence of functional epithelium. PLS regression analysis of H-1-NMR spectral data demonstrated that the spasmolytic activity was better correlated with signals for flavonol derivatives. Conclusions: Our data challenge the idea that warifteine and methylwarifteine mediate the spasmolytic activity of the polar extract of C. sympodialis leaves.