Yi-Yao Yang1, Zhao-Yu Wu1, Hao Zhang1, Shi-Jun Yin1, Fang-Bo Xia2, Qian Zhang1, Jian-Bo Wan2, Jian-Li Gao3, Feng-Qing Yang1. 1. 1School of Chemistry and Chemical Engineering, Chongqing University, Chongqing, 401331 People's Republic of China. 2. 3State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, People's Republic of China. 3. 2Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053 People's Republic of China.
Abstract
BACKGROUND: The dry root and rhizome of Salvia miltiorrhiza Bunge, or Danshen, is a well-known traditional Chinese medicine with anticoagulant activity. Taking into account that thrombin (THR) and factor Xa (FXa) play crucial roles in the coagulation cascade, it is reasonable and meaningful to screening THR and/or FXa inhibitors from Danshen. METHODS: Four extracts [butanol (BA), ethyl acetate (EA) and remained extract (RE) from 75% ethanol extract, and water extract (WE)] of Danshen were prepared, and their THR/FXa inhibitory activities were assessed in vitro. Then, the active EA extract was further separated by silica-gel column chromatography (SC), and its fractions (SC1-SC5) were analyzed by LC-MS. The principal component analysis (PCA) and orthogonal partial least squares discriminate analysis (OPLS-DA) were employed for predicting the specific marker compounds. The chemical structures of targeted compounds were identified by LC-MS/MS and their interactions with THR/FXa were analyzed by the molecular docking analysis. RESULTS: Danshen EA extract showed strong activity against THR and FXa, and its fractions (SC1-SC5) exhibited obvious difference in inhibitory activity against these two enzymes. Furthermore, four marker compounds with potential THR/FXa inhibitory activity were screened by PCA and OPLS-DA, and were identified as cryptotanshinone, tanshinone I, dihydrotanshinone I and tanshinone IIA. The molecular docking study showed that all these four tanshinones can interact with some key amino acid residues of the THR/FXa active cavities, such as HIS57 and SER195, which were considered to be promising candidates targeting THR and/or FXa with low binding energy (< - 7 kcal mol-1). CONCLUSIONS: LC-MS combined with multivariate statistical analysis can effectively screen potential THR/FXa inhibitory components in Danshen.
BACKGROUND: The dry root and rhizome of Salvia miltiorrhiza Bunge, or Danshen, is a well-known traditional Chinese medicine with anticoagulant activity. Taking into account that thrombin (THR) and factor Xa (FXa) play crucial roles in the coagulation cascade, it is reasonable and meaningful to screening THR and/or FXa inhibitors from Danshen. METHODS: Four extracts [butanol (BA), ethyl acetate (EA) and remained extract (RE) from 75% ethanol extract, and water extract (WE)] of Danshen were prepared, and their THR/FXa inhibitory activities were assessed in vitro. Then, the active EA extract was further separated by silica-gel column chromatography (SC), and its fractions (SC1-SC5) were analyzed by LC-MS. The principal component analysis (PCA) and orthogonal partial least squares discriminate analysis (OPLS-DA) were employed for predicting the specific marker compounds. The chemical structures of targeted compounds were identified by LC-MS/MS and their interactions with THR/FXa were analyzed by the molecular docking analysis. RESULTS: Danshen EA extract showed strong activity against THR and FXa, and its fractions (SC1-SC5) exhibited obvious difference in inhibitory activity against these two enzymes. Furthermore, four marker compounds with potential THR/FXa inhibitory activity were screened by PCA and OPLS-DA, and were identified as cryptotanshinone, tanshinone I, dihydrotanshinone I and tanshinone IIA. The molecular docking study showed that all these four tanshinones can interact with some key amino acid residues of the THR/FXa active cavities, such as HIS57 and SER195, which were considered to be promising candidates targeting THR and/or FXa with low binding energy (< - 7 kcal mol-1). CONCLUSIONS: LC-MS combined with multivariate statistical analysis can effectively screen potential THR/FXa inhibitory components in Danshen.
Authors: Robin J Rosenfeld; David S Goodsell; Rabi A Musah; Garrett M Morris; David B Goodin; Arthur J Olson Journal: J Comput Aided Mol Des Date: 2003-08 Impact factor: 3.686
Authors: Yi Luo; Yu Feng; Lei Song; Gan-Qing He; Sha Li; Sha-Sha Bai; Yu-Jie Huang; Si-Ying Li; Mohammed M Almutairi; Hong-Lian Shi; Qi Wang; Ming Hong Journal: Chin Med Date: 2019-08-06 Impact factor: 5.455