Synergy Mechanisms of Rhizoma Paridis Saponins on Non-small Cell Lung Cancer: Segmented Solid Phase Extraction, Bioactivity Screening, and Network Pharmacology.

BACKGROUND: Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. Rhizoma paridis saponins (RPS), the main bioactive ingredients of Paris polyphylla Smith var. yunnanensis (PPY), have been proved to have remarkable effects on NSCLC cell lines. However, the multi-component synergistic effects and mechanisms of RPS on NSCLC have not been elucidated. OBJECTIVE: To decipher the multi-RPS synergistic effects and mechanisms against NSCLC based on network pharmacology combined with segmented solid-phase extraction (SPE) and bioactivity screening method. METHODS: Firstly, segmented SPE and cytotoxicity assays were performed to screen the RPS-enrichment fraction of PPY, and the steroidal saponins in it were identified by LC-MS/MS. Then, a network pharmacology analysis was performed to predict the potential therapeutic targets of RPS on NSCLC. Finally, viable cell counting tests and RT-qPCR were utilized to verify the synergistic effects and mechanisms of RPS. RESULTS: 48 potentially active compounds were identified from the 30% MeOH/EtOAc fraction of PPY (30% M/E PPY). The results of the network pharmacology analysis indicated that RPS exerted joint effects by regulating six key targets in the PI3K-AKT signaling pathway. In vitro experiments showed that due to the synergistic effects, 30% M/E PPY at 13.90 µg/mL could exert a stronger inhibitory activity on A549 cells by reducing the overexpression of six hub genes compared with the parallel control groups. CONCLUSION: This research elaborates on the multi-RPS synergy mechanisms against NSCLC and provides a way to develop new combination medicines for NSCLC.

Rapid screening of neuraminidase inhibitors with the benzoic acid skeleton from Paeonia suffruticosa Andrews by solid-phase extraction with an enzyme activity switch combined with mass spectrometry analysis.

It is meaningful for drug discovery to discover lead compounds with specific skeletons from medicinal herbs. Screening bioactive compounds with specific skeletons by a simple and rapid strategy is still a challenging task. Solid-phase extraction (SPE) is a simple and time-saving technique in the laboratory and is often used in the concentration of natural products. It is attractive to apply the SPE in the screening of bioactive compounds with specific skeletons. To achieve this goal, SPE with an enzyme activity (EA) switch combined with mass spectrometry analysis was first proposed. The screening of benzoic acid-derived neuraminidase (NA) inhibitors from the root cortex of Paeonia suffruticosa Andrews (CPSA) was used as an example. The NA and crude extract of CPSA were incubated to form a sample solution. Subsequently, the sample was separated, detected, and collected by the SPE with an EA switch. When the detected values reduced significantly, the EA switch was triggered, and the collection was stopped. The collected eluents were treated for LC-MS/MS analysis. Finally, combining diagnostic ions and mass spectrometry data, two benzoic acid NA inhibitors were successfully screened from CPSA. In this study, the separation, detection, and collection were performed on one instrument system. Compared with the traditional isolation strategy, this strategy with the simpler operation and higher experimental efficiency could be an effective tool for the rapid screening of lead compounds with specific skeletons.