Zhongshan Zhongzhi Pharmaceutical Group Co., Ltd.

To investigate the chemical constituents of Shiqi Waigan Granules and their metabolic distribution in rats.

An off-line two-dimensional liquid chromatography-mass spectrometry approach was employed to identify the constituents of Shiqi Waigan Granules. A UPLC-QTOF-MS method was established to analyze the distribution of prototype compounds and metabolites in rat plasma, heart, liver, lung, kidney, spleen and brain after oral administration, and to elucidate the metabolic pathways.

(1) A total of 315 compounds were successfully identified in Shiqi Waigan Granules. Compared with conventional one-dimensional LC, the off-line 2D system amplified trace compounds and resolved more isomers, enabling a more systematic and refined characterization of the chemical profile. (2) Based on plasma pharmacochemistry and Phase I and II metabolic pathways, 66 prototype compounds and 102 metabolites were tentatively identified in rat plasma. (3) Most constituents were detected in liver, kidney, lung and spleen, whereas fewer were found in heart and brain.

Shiqi Waigan Granules are mainly composed of flavonoids, terpenoids, organic acids, lignans, phenolics, phenylpropanoids and glycosides. In vivo, these constituents primarily undergo hydrolysis, reduction, hydroxylation, carboxylation, sulfation, glucuronidation and composite reactions, and are widely distributed as either prototype compounds or their Phase I/II metabolites.

Phytochemical study of the n-BuOH extract of Ilex asprella resulted in the discovery of ten new pentacyclic triterpenoid glycosides, comprising nine ursane-type glycosides (1−9) and one oleanane-type glycoside (10), along with seven known compounds (11−17). Compound 1 is the first reported 19,22-epoxy ursane triterpenoid glycoside, whereas 4 and 5 are rare examples of ursane triterpenoid glycosides containing a 28,19-lactone group. The structural characterization of these compounds was achieved using spectroscopic and chemical techniques, as well as single-crystal X-ray analysis. Compounds 7, 12, 15, and 17 exhibited moderate cytotoxic activities against H1975 and HCC827 cancer cells.Graphical abstract

To dissect the therapeutic effect and potential pharmacological mechanisms of Keke tablets (KKT) against post-infectious cough (PIC) following influenza A virus infection based on network pharmacology, molecular docking, molecular dynamics, and in vivo experiments.

The chemical profiling of KKT was systematically analyzed using ultra-high performance liquid chromatography coupled with mass spectrometry (UPLC-MS). Active ingredients and targets of KKT were then screened through the TCMSP database, while disease-related targets of PIC were collected through the GeneCards database. Common core targets were obtained by drawing a Venn diagram, which were imported to the STRING database for network topology analysis. In addition, protein-protein interaction (PPI) network, Gene Ontology (GO) for core targets, functional analysis and pathway enrichment analysis from Kyoto Encyclopedia of Genes and Genomes (KEGG) were constructed to predict the network of KKT core active ingredients and PIC core targets. Molecular docking and dynamics analysis were performed to evaluate the interactions and movements of the components of KKT with the potential targets of PIC. A PIC model of guinea pigs was established by intranasally administering A/Puerto Rico/8/1934 H1N1/34 (A/PR/8) to verify the efficacy and potential pharmacological mechanism of KKT in treating PIC.

A total of 29 potential active ingredients and 602 related targets in KKT were screened following UPLC-MS analysis. After intersection with 718 potential targets in PIC, 10 core targets were identified through PPI analysis. GO and KEGG analyses indicated that KKT were mainly related to the inflammatory response, NOD-like receptor signaling pathway and Toll-like receptor signaling pathway in the treatment of PIC. Subsequently, the findings of network pharmacology were corroborated by molecular docking and molecular dynamics analysis, suggesting that the main active components in KKT may exert therapeutic effects on PIC by modulating the TLR4/NF-κB/NLRP3 signaling. In vivo experiments demonstrated that KKT reduced cough frequency of A/PR/8-challenged guinea pigs and inhibited the activation of TLR4/NF-κB/NLRP3 signaling pathway in lung tissues, implying that KKT could alleviate PIC by regulating TLR4/ NF-κB/NLRP3 signaling pathway and suppressing pyroptosis.

KKT could inhibit the activation of TLR4/NF-κB/NLRP3 signaling pathway to suppress the assembly of NLRP3 inflammasome and the secretion of cytokines and cough-related neuropeptides in treating influenza A virus-induced PIC.