Metabolomics-based comparative analysis of the effects of host and environment on Viscum coloratum metabolites and antioxidative activities.

Viscum coloratum (Kom.) Nakai is a well-known medicinal hemiparasite widely distributed in Asia. The synthesis and accumulation of its metabolites are affected by both environmental factors and the host plants, while the latter of which is usually overlooked. The purpose of this study was to comprehensively evaluate the effects of host and habitat on the metabolites in V. coloratum through multiple chemical and biological approaches. The metabolite profile of V. coloratum harvested from three different host plants in two habitats were determined by multiple chemical methods including high-performance liquid chromatography-ultraviolet (HPLC-UV), gas chromatography-flame ionization detector (GC-FID) and ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF/MS). The differences in antioxidant efficacy of V. coloratum were determined based on multiple in vitro models. The multivariate statistical analysis and data fusion strategy were applied to analyze the differences in metabolite profile and antioxidant activity of V. coloratum. Results indicated that the metabolite profile obtained by various chemical approaches was simultaneously affected by host and environment factors, and the environment plays a key role. Meanwhile, three main differential metabolites between two environment groups were identified. The results of antioxidant assay indicated that the environment has greater effects on the biological activity of V. coloratum than the host. Therefore, we conclude that the integration of various chemical and biological approaches combined with multivariate statistical and data fusion analysis, which can determine the influences of host plant and habitat on the metabolites, is a powerful strategy to control the quality of semi-parasitic herbal medicine.

Optimised architecture-based grading system as an independent prognostic factor in resected lung adenocarcinoma.

AIMS: Considering morphological heterogeneity of lung adenocarcinoma (LUAD) and no objective prognostic grading system existing currently, we aim to establish an 'optimised architecture-based grading system' (OAGS) to predict prognosis for resected LUAD. METHODS: A multicentral study involving three independent cohorts of LUAD was conducted. Predictive ability of the OAGS for recurrence-free probability (RFP) and overall survival (OS) was assessed in training cohort (n=228) by the area under the receiver operating characteristic curve (AUC), Harrell's concordance index (C-index) and Kaplan-Meier survival analyses, which was validated in testing (n=135) and validation (n=226) cohorts. RESULTS: The OAGS consists of: grade 1 for lepidic, papillary or acinar predominant tumour with no or less than 5% of high-grade patterns (cribriform, solid and or micropapillary), grade 2 for lepidic, papillary or acinar predominant tumour with 5% or more of high-grade patterns, and grade 3 for cribriform, solid or micropapillary predominant tumour. In all stages, the OAGS outperformed the pattern-dominant grading system and IASLC grading system for predicting RFP (C-index, 0.649; AUC, 0.742) and OS (C-index, 0.685; AUC, 0.754). Multivariate analysis identified it as an independent predictor of both (RFP, p<0.001; OS, p<0.001). Furthermore, in pT1-2aN0M0 subgroup, the OAGS maintained its ability to predict recurrence (C-index, 0.699; AUC, 0.769) and stratified patients into different risk groups of RFP (p<0.001). These results were confirmed in testing and validation cohorts. CONCLUSIONS: The OAGS is an independent prognostic factor and shows a robust ability to predict prognosis for resected LUAD.

Exploring stereoselective excretion and metabolism studies of novel 2-(2-hydroxypropanamido)-5-trifluoromethyl benzoic acid enantiomers.

R-/S-2-(2-Hydroxypropanamido)-5-trifluoromethyl benzoic acid (R-/S-HFBA), as a novel COX inhibitor, was firstly reported to have remarkable anti-inflammatory and antiplatelet aggregation activities by our group. In our previous study, stereoselective differences in pharmacokinetics were found between HFBA enantiomers after oral and intravenous administration of each enantiomer to rats. The discrepancies might be associated with the excretion and metabolism of the two enantiomers. In this research, an UHPLC-MS/MS method was established and validated for quantification of R-/S-HFBA in rats urine, feces and bile samples in excretion study. Moreover, an ultra high performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) method was employed to understand the metabolism of R-/S-HFBA in rats. Results showed that the total cumulative excretion of R-/S-HFBA in three routes were 65.8% and 58.5% of the dose, respectively. The urinary excretion of R-/S-HFBA was the main route, which accounted for 40.2% and 31.7% respectively; the cumulative biliary excretion of R-/S-HFBA were 11.3% and 7.4%; the cumulative amounts of R-/S-HFBA excreted directly via feces without absorption from the gastrointestinal tract were 14.3% and 19.4%, respectively. R-/S-HFBA existed stereoselective discrepancy in excretion. In addition, 8 metabolites of S-HFBA were detected and tentatively identified including glucuronidation, glycine and N-acetyl conjugation while R-HFBA existed 7 metabolites without glycine conjugation. Formation of metabolites of R-/S-HFBA also exhibited stereoselectivity. In summary, these new findings on excretion and metabolism of R-/S-HFBA provided valuable information for stereoselective pharmacokinetics and were greatly helpful for further investigation, such as safety and mechanism of action.

Synthesis, pharmacological evaluation and molecular docking of novel R-/S-2-(2-hydroxypropanamido)-5-trifluoromethyl benzoic acid as dual anti-inflammatory anti-platelet aggregation agents.

R-/S-2-(2-hydroxypropanamido) benzoic acid (R-/S-HPABA), marine-derived anti-inflammatory antiplatelet drugs, were initially synthesised in our group. However, preliminary research showed that R-/S-HPABA were eliminated rapidly because of extensive hydroxylation metabolism of phenyl ring in vivo. In order to reduce significant hydroxylation metabolism to improve pharmacological activity and bioavailability, trifluoromethyl group was incorporated into R-/S-HPABA to synthesise R-/S-2-(2-hydroxypropanamido)-5-trifluoromethyl benzoic acid (R-/S-HFBA), respectively. The purposes of this study were to report the synthesis of R-/S-HFBA and compare the anti-inflammatory antiplatelet effect and pharmacokinetic properties of R-/S-HFBA with those of R-/S-HPABA. Carrageenan-induced rat paw edema assay was used for the evaluation of the anti-inflammatory activity. R-/S-HFBA showed better results in inhibiting edema and were able to prolong the anti-inflammatory effect after carrageenan injection. The antiplatelet aggregation activity of R-/S-HFBA and R-/S-HPABA was studied on arachidonic acid-induced platelet aggregation of rabbit platelet-rich plasma. The aggregation inhibition rate of R-/S-HFBA was significantly (p < 0.05) higher than that of R-/S-HPABA, respectively. Molecular docking study revealed that R-/S-HFBA possess more potent binding affinity with COX-1/COX-2 than R-/S-HPABA, respectively, and that the presence of trifluoromethyl group leads to increase in activity of R-/S-HFBA. R-/S-HFBA also afford more favorable pharmacokinetic properties than R-/S-HPABA, respectively, such as higher Cmax, larger AUC0-∞, and longer t1/2, which, as expected, are more metabolically stable.