A multidimensional strategy for characterization, distinction, and quality control of two Clinopodium medicinal plants.

ETHNOPHARMACOLOGICAL RELEVANCE: Clinopodium chinense Kuntze (CC) and Clinopodium polycephalum (Vaniot) C. Y. Wu & S. J. Hsuan (CP) are both included in the Pharmacopoeia of the People's Republic of China (edition 2020) as the legitimate source of "Duan Xue Liu" (DXL), which is a crucial traditional Chinese medicine used as a clinical remedy for bleeding diseases. However, the differences in plant endogenous metabolites and bioactivities between CC and CP are still unclear. AIM OF THE STUDY: This study aims to provide a scientific basis to investigate the differences between CC and CP ensuring the efficient and safe use of DXL. MATERIALS AND METHODS: A multidimensional strategy including plant metabolomics, digital reference standard (DRS) analyzer, and biological activities assay was creatively constructed for the characterization, distinction, and quality control of CC and CP. RESULTS: There were apparent differences in the metabolites between CC and CP. 7 compounds contributing to the differences were successfully identified. On that basis, linear calibration using two reference substances (LCTRS) methods was proved as a more accurate and specific quality analysis method for CC and CP. In addition, bioactivity assays showed that both CC and CP exhibited obvious hemostatic activity, while CC showed greater potential to resist inflammation and free radicals. CONCLUSION: In summary, it was the first time to investigate the chemical constituents and bioactivities differences between CC and CP with the help of plant metabolomics, DRS study, and biological activity assays. These two plants were significantly separated in the integrated analysis, suggesting that we should pay attention to the distinction to prevent unexpected risks caused by medicinal materials.

Monoterpenoid indole alkaloids from Melodinus axillaris W.T.Wang exhibit anti-inflammatory activities by inhibiting the NF-κB signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Melodinus axillaris W.T.Wang has been widely used as an important medicine in China. In the folk of China, its whole plant has been used for fractures, rheumatic heart disease, testitis, hernia, abdominal pain, and dyspepsia, etc. Despite its extensive use, there is a shortage of literature investigating the specific bioactive compounds and underlying mechanisms responsible for their anti-inflammatory effects. This knowledge gap serves as the primary impetus for conducting this study, which aims to shed light on the previously unexplored therapeutic potential of M. axillaris. AIM OF THE STUDY: This study aims to investigate the material basis and potential mechanism of anti-inflammatory activity of M. axillaris. MATERIALS AND METHODS: Compounds were isolated from the 95% ethanol extract of M. axillaris using a systematic phytochemical method. The structures were established by extensive spectroscopic analysis, including 1D and 2D NMR, HR-ESI-MS, ECD calculation, and DP4+ analysis. The anti-inflammatory activities of ethanol extract and compounds from M. axillaris were tested by an inflammation model of LPS-stimulated RAW264.7 cells in vitro. Western blot analysis was employed to evaluate the expressions of COX-2, iNOS, and NF-κB signaling pathways, aiming to elucidate the underlying mechanisms. RESULTS: Eleven undescribed monoterpenoid indole alkaloids (MIAs), axillines A-K (1-11), along with thirteen known analogs were isolated from M. axillaris. Compound 1 was the first representative of vincadine alkaloid with unprecedented 6/5/9/6/6 skeletons. Compounds 1-11 and ethanol extract showed significant anti-inflammatory effects in vitro. Among them, compound 2 had the best activity of inhibiting NO release (IC50 = 3.7 ± 0.9 µM). Additionally, subsequent Western blot analysis revealed that 2 could significantly inhibit the up-regulation of NF-κB signaling pathways, iNOS, and COX-2 in LPS-stimulated RAW264.7 cells, thereby demonstrating its anti-inflammatory activity. CONCLUSION: This study provides support for the traditional use of M. axillaris in terms of its anti-inflammatory properties and highlights the potential of MIAs as promising candidates for further development as anti-inflammatory drugs.

Monoterpenoid indole alkaloid dimers from the Melodinus axillaris induce G2/M phase arrest and apoptosis via p38 MAPK activation in HCT116 cells.

Four monoterpenoid indole alkaloid dimers (MIADs), axidimins A-D (1-4), which possesses unprecedented apidosperma-aspidosperma-type skeletons, along with twelve known MIAs were isolated from Melodinus axillaris. Their structures were established by comprehensive analysis of the HRESIMS, NMR, ECD calculation and DP4 + analysis. A possible biosynthetic pathway for axidimins A-D was proposed. In vitro, axidimins C and D exhibited significant cytotoxicities against HCT116 cells with IC50 values of 5.3 µM and 3.9 µM, respectively. The results obtained from flow cytometry and Western blot analysis clearly demonstrated that axidimins C and D significantly induced a reverse G2/M phase arrest and apoptosis of HCT116 cells. The potential mechanism of axidimins C and D on HCT116 cells were thoroughly discussed through the utilization of network pharmacology and molecular docking research. Subsequently, the selected targets were validated using Western blot and CETSA analysis, confirming that axidimins C and D exert its cytotoxic effects through the activation of the p38 MAPK pathway, ultimately leading to HCT116 cells death. This study provides evidence indicating that axidimins C and D have the potential to induce cell cycle arrest and apoptosis in HCT116 cells by modulating the p38 MAPK signaling pathway. These findings offer a novel perspective for the development of anti-colorectal cancer drugs.

Kamaonensine A-G: Lycaconitine-type C19-diterpenoid alkaloids with anti-inflammatory activities from Delphinium kamaonense Huth.

Delphinium kamaonense Huth is a sort of folkloric plant resource which is cultivated and planted with great ornamental and medicinal values. In this work, seven undescribed lycaconitine-type C19-diterpenoid alkaloids, especially a rare skeleton with -CH=N and N-oxide moieties, along with ten known compounds, were isolated from D. kamaonense, of which the structures were determined by various spectroscopic data, combined with calculated electronic circular dichroism (ECD) and single-crystal X-ray diffraction analysis. In vitro nitric oxide inhibitory activities assay of these compounds indicated that lycaconitine-type C19-diterpenoid alkaloids had significant anti-inflammatory inhibitory activities, with kamaonensine E being the most potent (0.9 ± 0.2 µM) stronger than positive (9.0 ± 1.3 µM). In the network pharmacology studies, binding three key targets mitogen-activated protein kinase 8 (MAPK8), mitogen-activated protein kinase 14 (MAPK14), and heat shock protein HSP 90-alpha (HSP90α), the anti-inflammatory mechanism might be related to MAPK signaling pathways. Furthermore, the molecular docking results revealed that the uncommon amides and methylenedioxy groups might be the most two promising pharmacophores for lycaconitine-type C19-diterpenoid alkaloids.

Three Undescribed Flavonoids and Amide from Abrus mollis Hance with Hepatoprotective Effects.

Through a phytochemical investigation of Abrus mollis Hance, a folk medicinal plant in China, we isolated and identified three undescribed compounds, including two flavonoids and one amides alkaloid, along with nine known from this plant. Their structures were elucidated by analyses of 1D, 2D NMR, HR-ESI-MS, ECD, and DP4+ analysis. Furthermore, we evaluated the hepatoprotective effects of all twelve compounds on D-GalN-induced Brl-3 A cells. According to the results, at a concentration of 25 µM, the cell survival rates were observed to be 71.92±0.34 %, 70.03±1.29 %, and 69.11±1.90 % for compound 2, 4, and 11, respectively. Further experimental studies showed that compound 2 (EC50 5.76±0.37 µM) showed more significant protective activity than the bicyclol.

Diterpene glycosides, acetophenone glycosides and tannins from polar extracts of the root of Euphorbia fischeriana with cytotoxicity and antibacterial activities.

The phytochemical study on Euphorbia fischeriana, a folk medicinal plant in China, led to the isolation of eight undescribed glycosides, including two diterpene glycosides, three acetophenone glycosides and three tannins together with eight known ones. Their planar structures were elucidated by extensive analyses of 1D, 2D NMR experiments and HRESIMS. The absolute configurations were determined by NOESY experiments, ECD calculations. All undescribed compounds were evaluated for their cytotoxicity and antibacterial activities in vitro. Two diterpene glycosides (1-2) showed cytotoxic activity with IC50 values ranging from 5.4 to 16.2 µM toward Hep-G2, Hep-3B, A549, NCI-H460 and AGS cells. Tannins (6-8) showed the significant antibacterial activity with MIC values in the range of 1.56-6.25 µg/mL.

Isoquinoline alkaloids from the Corydalis tomentella with potential anti-hepatoma and antibacterial activities.

Six undescribed isoquinolines, including one rarely reported N-benzyl isoquinoline together with sixteen known ones were isolated from C. tomentella. Their planar structures and absolute configurations were elucidated by extensive analyses of UV, IR, NMR, HRESIMS, DP4+ probability analysis as well as ECD calculations. Biological evaluations revealed that 3,4-2H-tomentelline C (6) showed significant cytotoxicity (IC50 = 7.42 µM) against the HepG2 cell line while (1'R, 2'S)-coptichine B (3) exhibited stronger antibacterial activities.

Secondary Metabolites from the Fresh Leaves of Pinus yunnanensis Franch.

Fifteen metabolites, including two flavonols (1-2), three lignans (3-5), and ten diterpenoids (6-15), were isolated from the leaves of Pinus yunnanensis. Among them, flavanonol (1) were identified as undescribed flavonol derivative with natural rarely B-ring fission lactone. Massive spectroscopic methods, the DP4+ probabilities and CD/ECD calculations were applied to establish the structure of component 1. Among these compounds, taxifolin (2) showed potent cytotoxicity, having IC50 values from 21.33 to 45.48 µg/mL, it also showed broad antibacterial activity against human pathogens with MIC values from 32 to 64 µg/mL.

Isolation of triterpenoids and phytosterones from Achyranthes bidentata Bl. to treat breast cancer based on network pharmacology.

Breast cancer (BC) is a cancer with a high incidence and mortality of female, threatening women's physical and mental health. Achyranthes bidentata Blume, a traditional Chinese medicine, has been popular in folk due to its effective pharmacology activities including anti-inflammatory and anti-tumour effects. In order to identify the active ingredients from the roots of A. bidentata for treating breast cancer, the separation guided by network pharmacology analysis was employed which led to the isolation of 7 triterpenoids and 10 phytosterones. According to the in vitro experiments, the isolated compounds showed significant inhibitory activities on MCF-7 cells as well as anti-inflammatory activities by attenuating the production of NO and TNF-α in LPS-induced macrophage cells. In conclusion, this study successfully predicted and confirmed the positive impact of triterpenoids and phytosterones on breast cancer, which provided a foundation for further in vivo studies.

Antiproliferative diterpenoids and acetophenone glycoside from the roots of Euphorbia fischeriana.

Three undescribed compounds and nine known compounds were isolated from the Euphorbia fischeriana Steud. Their structures were elucidated by extensive analysis of the 1D and 2D NMR spectra and HRESIMS data. The absolute configurations of two undescribed diterpenoids were determined by comparing their experimental and calculated ECD data. To further understand the antitumor effects of E. fischeriana, we tested the antiproliferative activities of these compounds against AGS, Hep-G2 cell lines in vitro using CCK-8 assays. The results showed that both types of compounds had significant antiproliferative effects, among which a diterpenoid with a galloyl moiety (IC50 = 7.03 µM) was the most significant one that could be regarded as a potential leading compound against liver cancer for further comprehensive study.

Polyhydroxyl guaianolide terpenoids as potential NF-кB inhibitors induced cytotoxicity in human gastric adenocarcinoma cell line.

Six new guaiane-type sesquiterpenes (1-6), and one monoterpenoid (7) along with five known analogues (8-12), were isolated from the leaves of Artemisia argyi Lévl et Vant. The new compounds were characterized by the basic analysis of the spectroscopic data (HRMS, 1D and 2D NMR), and the absolute configurations were determined by both calculated electronic circular dichroism and DP4 calculations. The inhibitory effects of 1-12 against human gastric adenocarcinoma (AGS) cells were investigated in vitro, among which 1-3 and 8 showed remarkable cytotoxic activity with IC50 values in the range of 6.69-10.25 µM. The results suggested that the variation in the inhibitory activities of the compounds are the result of different substitutions on C-8. In order to rationalize the binding interactions of active compounds with the active site of NF-кB, in silico study was conducted and the results were in complete agreement with the experimental data for cytotoxicity evaluation.

New nor-oleanane triterpenoids from the fruits of Stauntonia brachyanthera with potential anti-inflammation activity.

The continuing studies on the fruits of Stauntonia brachyanthera led to the isolations of 6 normal and nor-oleanane triterpenoids, including two new compounds, brachyantheoraside B10 (1) and brachyantheraside C1 (2). Compounds 1-3 showed inhibitory activity on lipopolysaccharide (LPS)-induced nitric oxide production in RAW 264.7 macrophages with the IC50 values of 27.04, 26.22, and 21.41 µM, respectively, which were all stronger than that of indomethacin. The study not only revealed a new type of nor-oleanane triterpenoids, but also confirmed the anti-inflammation abilities of three type nor-oleanane triterpenoids as the inhibitors of NO production.

Systems Pharmacological Approach to Investigate the Mechanism of Ohwia caudata for Application to Alzheimer's Disease.

Ohwia caudata (OC)-a traditional Chinese medicine (TCM)-has been reported to have large numbers of flavonoids, alkaloids, and triterpenoids. The previous studies on OC for treating Alzheimer's disease (AD) only focused on single targets and its mechanisms, while no report had shown about the synergistic mechanism of the constituents from OC related to their potential treatment on dementia in any database. This study aimed to predict the bioactive targets constituents and find potential compounds from OC with better oral bioavailability and blood-brain barrier permeability against AD, by using a system network level-based in silico approach. The results revealed that two new flavonoids, and another 26 compounds isolated from OC in our lab, were highly connected to AD-related signaling pathways and biological processes, which were confirmed by compound-target network, Gene Ontology (GO) analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, respectively. Predicted by the virtual screening and various network pharmacology methods, we found the multiple mechanisms of OC, which are effective for alleviating AD symptoms through multiple targets in a synergetic way.