Withanolides from Physalis angulata var. villosa and the Relative Configurational Revision of Some Known Analogs.

Physalis angulata var. villosa is a plant possessing abundant withanolides, but in-depth research is lacking. In our ongoing study of P. angulata var. villosa, 15 previously undescribed withanolides (1-15), along with 21 known analogs (16-36), were isolated from the whole plant. The structures of the withanolides (1-15) were elucidated based on analysis of their 1D and 2D NMR, HRESIMS, and ECD data. Additionally, the application of γ-gauche effects with the help of ROESY correlations led to the formulation of empirical rules for withanolides with 14-OH/15-OAc to rapidly determine the 14-OH orientations, making it possible to propose configurational revisions of 19 previously reported analogs (1'-19'). Withanolides 1, 4-6, and 10 showed potent cytotoxic activities against three human cancer cell lines (HCT-116, MDA-MB-231, and A549).

Blood cell for the differentiation of airway inflammatory phenotypes in COPD exacerbations.

BACKGROUND: Measurement of sputum is frequently used to define airway inflammatory subtypes. The venous blood cell is a reliable and simple biomarker, may be used as an alternative procedure to reflect the subtypes. For the aim of verifying the hypothesis that venous blood cell can quantify sputum inflammatory cell to access the airway subtypes in chronic obstructive pulmonary disease of acute exacerbations (AECOPD) and to ascertain the accuracy of the blood cell biomarker. METHODS: This study evaluated 287 patients with COPD exacerbations and all four tests were performed on the same day, which are lung function test, bronchodilator reversibility test, sputum cell analysis and blood routine examination. RESULTS: There was a correlation between sputum eosinophils and blood eosinophils, blood cells derived ratios. There was a weaker relationship to neutrophils between sputum and blood. Sputum neutrophils had not any association with neutrophil/macrophage ratio (NMR) and eosinophil/lymphocyte ratio (ELR) in blood. Blood eosinophils percentage was predictive for eosinophilic COPD exacerbations with an area under the curve (AUC) of 0.672 (p = 0.012). The optimum cutpoint for blood eosinophils percentage was 0.55%. Blood eosinophils absolute count was also predictive sputum eosinophilia at 0.35 × 109/L (AUC = 0.626, p = 0.025). ELR, eosinophil/monocyte ratio (EMR) and eosinophil/neutrophil ratio (ENR) in blood were higher in COPD exacerbations with mixed granulocytic and eosinophilic subtypes. CONCLUSION: Eosinophils/neutrophils count parameters were relationship between blood and sputum. Eosinophils in blood and the ratios (ENR, EMR and ELR) may be utilized to assess eosinophilic airway inflammation in COPD exacerbations. Due to weak relationship and poor predictive ability, more researches should be required.

Paper-based ligand fishing method for rapid screening and real-time capturing of α-glucosidase inhibitors from the Chinese herbs.

Identifying medicinally relevant compounds from natural resources generally involves the tedious work of screening plants for the desired activity before capturing the bioactive molecules from them. In this work, we created a paper-based ligand fishing platform to vastly simplify the discovery process. This paper-based method exploits the enzymatic cascade reaction between α-glucosidase (GAA), glucose oxidase (GOx), and horseradish peroxidase (HRP), to simultaneously screen the plants and capture the GAA inhibitors from them. The designed test strip could capture ligands in tandem with screening the plants, and it features a very simply operation based on direct visual assessment. Multiple acylated flavonol glycosides from the leaves of Quercus variabilis Blume were newly found to possess GAA inhibitory activities, and they may be potential leads for new antidiabetic medications. Our study demonstrates the prospect of the newly discovered GAA ligands as potential bioactive ingredients as well as the utility of the paper-based ligand fishing method.

New N-oxide alkaloids from the stems of Sinomenium acutum.

Six new alkaloids (1-6) and six known alkaloids (7-12) were obtained from the stems of Sinomenium acutum. Among them, compounds 1-3 and 6 were four N-oxide alkaloids. The structures and absolute configurations of these new alkaloids were elucidated through comprehensive data of 1D and 2D NMR, HRESIMS and ECD spectra. All isolated compounds were evaluated in vitro for their inhibitory activities against nitric oxide (NO) production and inhibitory effects on AChE. Among them, the sinomenine N-oxide (9) was the most potent NO production inhibitor, with an IC50 value of 23.04 µM.

Identification of a novel PHGDH covalent inhibitor by chemical proteomics and phenotypic profiling.

The first rate-limiting enzyme of the serine synthesis pathway (SSP), phosphoglycerate dehydrogenase (PHGDH), is hyperactive in multiple tumors, which leads to the activation of SSP and promotes tumorigenesis. However, only a few inhibitors of PHGDH have been discovered to date, especially the covalent inhibitors of PHGDH. Here, we identified withangulatin A (WA), a natural small molecule, as a novel covalent inhibitor of PHGDH. Affinity-based protein profiling identified that WA could directly bind to PHGDH and inactivate the enzyme activity of PHGDH. Biolayer interferometry and LC-MS/MS analysis further demonstrated the selective covalent binding of WA to the cysteine 295 residue (Cys295) of PHGDH. With the covalent modification of Cys295, WA blocked the substrate-binding domain (SBD) of PHGDH and exerted an allosteric effect to induce PHGDH inactivation. Further studies revealed that with the inhibition of PHGDH mediated by WA, the glutathione synthesis was decreased and intracellular levels of reactive oxygen species (ROS) were elevated, leading to the inhibition of tumor proliferation. This study indicates WA as a novel PHGDH covalent inhibitor, which identifies Cys295 as a novel allosteric regulatory site of PHGDH and holds great potential in developing anti-tumor agents for targeting PHGDH.

Inflammatory Subtypes in Classic Asthma and Cough Variant Asthma.

BACKGROUND: Measurement of sputum is used to define airway inflammatory phenotypes. Cough variant asthma (CVA) is considered to be the initial stage of classic asthma (CA). The aim of this study was to describe the association between the different subtypes of CVA and CA. METHODS: A total of 459 patients with CVA and CA were screened for the study. All included patients performed spirometry, underwent a bronchial challenge with methacholine and induced sputum according to the guidelines. RESULTS: A higher frequency of female patients were found with CVA and the eosinophilic airway inflammation of CVA than in CA and the noneosinophilic airway inflammation of CA (p=0.004 and p=0.024, respectively). Bronchial hyper-responsiveness (BHR) was lower in eosinophilic CVA and CA (p=0.006), while no difference was found in noneosinophilic CVA and CA. Association between the percentage of sputum eosinophils and the FEV1 level fell below 20% of the baseline value (PD20) in CVA and CA (r= -0.1245, p=0.0357 and r= -0.2148, p=0.0014, respectively). CONCLUSION: Eosinophilia may be associated with more severe disease, yet there was no difference in spirometry between the eosinophilic and noneosinophilic groups, and the BHR difference was not dramatic.

Sputum cell count: biomarkers in the differentiation of asthma, COPD and asthma-COPD overlap.

INTRODUCTION: Cell count in induced sputum is a noninvasive biomarker to assess airway inflammation phenotypes. Accordingly, sputum cell counts are extensively used in the treatment of asthma and COPD. Nevertheless, the clinical application of sputum cell counts in patients with asthma-COPD overlap (ACO) remains elusive. The aim of this study was to investigate sputum cell counts in patients with ACO which are different from those in patients with asthma and COPD and also to examine the relationship between sputum cell counts in bronchial reversibility and bronchial hyperresponsiveness (BHR). PATIENTS AND METHODS: A total of 374 patients participated in the study, including 142 patients with asthma, 160 patients with COPD and 72 patients with ACO. All patients underwent the following tests on the same day: pulmonary function test (PFT), BHR test or bronchodilator reversibility test and inducing sputum. They were classified into the asthma group, COPD group or ACO group based on a clinical history, PFT values and BHR test or bronchodilator reversibility test. RESULTS: The three groups had different PFT values (p<0.001) except for forced vital capacity (FVC) between the asthma and ACO groups (p=0.378). The sputum levels of eosinophil% were decreased in patients with COPD when compared with those in patients with asthma and ACO (p<0.001 and p<0.001, respectively). There was a difference in sputum neutrophil% and macrophage% counts among the three groups (p<0.001 and p<0.001, respectively); there was no difference in sputum eosinophil% counts between patients with ACO and asthma (p=0.668) and there was no difference in the percentage of induced sputum cells between the stage of airway obstruction and the stage of BHR. CONCLUSION: The clinical relevance of this study provides evidence that sputum cell counts as an inflammatory biomarker could carry some information to distinguish ACO, asthma and COPD, and these biomarkers need more studies to provide diagnostic value in the differentiation between ACO, asthma and COPD.