Dose-dependent targeted knockout methodology combined with deep structure elucidation strategies for Chinese licorice chemical profiling

et al. 2016

Sci Rep

Self Cite

The nucleoprotein (NP) of Ebola virus (EBOV) and Marburg virus (MARV) is an essential component of the viral ribonucleoprotein complex and significantly impacts replication and transcription of the viral RNA genome. Although NP is regarded as a promising antiviral druggable target, no chemical ligands have been reported to interact with EBOV NP or MARV NP. We identified two compounds from a traditional Chinese medicine Gancao (licorice root) that can bind both NPs by combining affinity mass spectrometry and metabolomics approaches. These two ligands, 18β-glycyrrhetinic acid and licochalcone A, were verified by defined compound mixture screens and further characterized with individual ligand binding assays. Accompanying biophysical analyses demonstrate that binding of 18β-glycyrrhetinic acid to EBOV NP significantly reduces protein thermal stability, induces formation of large NP oligomers, and disrupts the critical association of viral ssRNA with NP complexes whereas the compound showed no such activity on MARV NP. Our study has revealed the substantial potential of new analytical techniques in ligand discovery from natural herb resources. In addition, identification of a chemical ligand that influences the oligomeric state and RNA-binding function of EBOV NP sheds new light on antiviral drug development.

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Novel Chemical Ligands to Ebola Virus and Marburg Virus Nucleoproteins Identified by Combining Affinity Mass Spectrometry and Metabolomics Approaches

et al. 2016

Sci Rep

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“…Compounds 101 (t R = 15.48 min) and 104 (t R = 15.64 min) produced [M − H] − ions at m/z 591.1728 and 459.1305 and subsequently lost an acetyl group (C 2 H 2 O) to give a series of fragment ions similar to those of isoliquiritin apioside and liquiritin, respectively. Therefore, compounds 101 and 104 were tentatively identified as the reported acetyl isoliquiritin apioside [23] and 6″‐acetylliquiritin [24].…”

Section: Resultsmentioning

confidence: 99%

“…Among these, compounds O) to give a series of fragment ions similar to those of isoliquiritin apioside and liquiritin, respectively. Therefore, compounds 101 and 104 were tentatively identified as the reported acetyl isoliquiritin apioside [23] and 6″-acetylliquiritin [24]. ) and m/z 80.9652 (HSO 3 − ) were generated through the loss of the H 2 SO 3 moiety from the deprotonated ion.…”

Section: Flavanones Chalcones and Their Glycosidesmentioning

confidence: 99%

Deciphering the chemical constituents of Shengjiang Xiexin decoction by ultrahigh‐performance liquid chromatography‐quadrupole/orbitrap high‐resolution mass spectrometry and the impact of 20 characteristic components on multidrug resistance‐associated protein 2 in the vesicular transport assay

Guan

J of Separation Science

et al. 2022

Shengjiang Xiexin decoction, a traditional Chinese medical formula, has been utilized to alleviate the delayed‐onset diarrhea induced by irino tecan. However, the chemical constituents of this formula and the activities of its constituents remain unclear. In this study, ultrahigh‐performance liquid chromatography‐quadrupole/orbitrap high‐resolution mass spectrometry was employed to comprehensively analyze the chemical constituents of Shengjiang Xiexin decoction. A total of 270 components, including flavonoids, coumarins, triterpenoids, alkaloids, diarylheptanoids and others, were identified or characterized. Multidrug resistance‐associated protein 2 is an efflux transporter responsible for regulating drug absorption. A total of 20 characteristic components from the formula were selected to evaluate their effects on the function of multidrug resistance‐associated protein 2 using the vesicular transport assay. Glycyrrhizic acid and glycyrrhetinic acid were identified as potential multidrug resistance‐associated protein 2 inhibitors, while 9 flavonoid aglycones increased the uptake of the substrate [3H]‐estradiol 17‐β‐glucuronide in the vesicles. This was the first systematic investigation of the chemical constituents from Shengjiang Xiexin decoction and the effect of its characteristic components on the transporter. The results offered a basis for further exploring the detoxification mechanisms of this formula and its interactions with other drugs.

“…But the constituents in TCM are complex, and their concentrations in TCM differ by several orders of magnitude. Generally 50 to more than 100 constituents are detected in the whole extract of TCM formula by LC-MS 2 3 . Preliminary fractionation could enrich the concentrations of trace constituents, which would be helpful for detecting trace constituents in TCM formula by LC-MS analysis 4 5 .…”

mentioning

confidence: 99%

Rapid discovery and identification of anti-inflammatory constituents from traditional Chinese medicine formula by activity index, LC-MS, and NMR

Liu

et al. 2016

Sci Rep

The traditional activity-guided approach has the shortcoming of low accuracy and efficiency in discovering active compounds from TCM. In this work, an approach was developed by integrating activity index (AI), liquid chromatography – mass spectrometry (LC-MS), and nuclear magnetic resonance (NMR) to rapidly predict and identify the potential active constituents from TCM. This approach was used to discover and identify the anti-inflammatory constituents from a TCM formula, Gui-Zhi-Jia-Shao-Yao-Tang (GZJSYT). The AI results indicated that, among the 903 constituents detected in GZJSYT by LC-MS, 61 constituents with higher AI values were very likely to have anti-inflammatory activities. And eight potential active constituents of them were isolated and validated to have significant inhibitory effects against NO production on LPS-induced RAW 264.7 cell model. Among them, glycyrrhisoflavone (836), glisoflavanone (893) and isoangustone A (902) were reported to have anti-inflammatory effects for the first time. The proposed approach could be generally applicable for rapid and high efficient discovery of anti-inflammatory constituents from other TCM formulae or natural products.