Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Chinese Medicine
Published in: Chinese Medicine 1/2020

Open Access 01-12-2020 | Research

LC–MS-based multivariate statistical analysis for the screening of potential thrombin/factor Xa inhibitors from Radix Salvia Miltiorrhiza

Authors: Yi-Yao Yang, Zhao-Yu Wu, Hao Zhang, Shi-Jun Yin, Fang-Bo Xia, Qian Zhang, Jian-Bo Wan, Jian-Li Gao, Feng-Qing Yang

Published in: Chinese Medicine | Issue 1/2020

Login to get access

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.
Literature
1.
Zhong GX, Li P, Zeng LJ, Guan J, Li DQ, Li SP. Chemical characteristics of Salvia miltiorrhiza (Danshen) collected from different locations in China. J Agric Food Chem. 2009;57(15):6879–87.PubMedCrossRef
2.
Zhan ZL, Fang WT, Ma XH, Chen T, Cui GH, Ma Y, et al. Metabolome and transcriptome analyses reveal quality change in the orange-rooted Salvia miltiorrhiza (Danshen) from cultivated field. Chin Med. 2019;14:42.PubMedPubMedCentralCrossRef
3.
Luo Y, Feng Y, Song L, He GQ, Li S, Bai SS, et al. A network pharmacology-based study on the anti-hepatoma effect of Radix Salviae Miltiorrhizae. Chin Med. 2019;14:27.PubMedPubMedCentralCrossRef
4.
Luo H, Vong CT, Chen HB, Gao Y, Lyu P, Qiu L, et al. Naturally occurring anti-cancer compounds: shining from Chinese herbal medicine. Chin Med. 2019;14:18.CrossRef
5.
Gao CF, Liu LJ, Zhou YY, Bian ZX, Wang SP, Wang YT. Novel drug delivery systems of Chinese medicine for the treatment of inflammatory bowel disease. Chin Med. 2019;14:23.PubMedPubMedCentralCrossRef
6.
Chong CM, Su HX, Lu JJ, Wang YT. The effects of bioactive components from the rhizome of Salvia miltiorrhiza (Danshen) on the characteristics of Alzheimer’s disease. Chin Med. 2019;14:19.PubMedPubMedCentralCrossRef
7.
Kang DG, Yun YG, Ryoo JH, Lee HS. Anti-hypertensive effect of water extract of Danshen on renovascular hypertension through inhibition of the renin angiotensin system. Am J Chin Med. 2002;30(1):87–93.PubMedCrossRef
8.
Piao CL, Luo JL, Jin D, Tang C, Wang L, Lian FM, et al. Utilizing network pharmacology to explore the underlying mechanism of Radix Salviae in diabetic retinopathy. Chin Med. 2019;14:58.PubMedPubMedCentralCrossRef
9.
10.
Han JY, Fan JY, Horie Y, Miura S, Cui DH, Ishii H, et al. Ameliorating effects of compounds derived from Salvia miltiorrhiza root extract on microcirculatory disturbance and target organ injury by ischemia and reperfusion. Pharmacol Ther. 2008;117(2):280–95.PubMedCrossRef
11.
Lloyd-Jones DM, Wang TJ, Leip EP, Larson MG, Levy D, Vasan RS, et al. Lifetime risk for development of atrial fibrillation: the Framingham Heart Study. Circulation. 2004;110(9):1042–6.PubMedCrossRef
12.
Birrell F, Johnell O, Silman A. Projecting the need for hip replacement over the next three decades: influence of changing demography and threshold for surgery. Ann Rheum Dis. 1999;58(9):569–72.PubMedPubMedCentralCrossRef
13.
Fareed J, Jeske WP. Small-molecule direct antithrombins: argatroban. Best Pract Res Clin Haematol. 2004;17(1):127–38.PubMedCrossRef
14.
Wang WZ, Yuan J, Fu XL, Meng FC, Zhang SJ, Xu WR, et al. Novel anthranilamide-based FXa inhibitors: drug design, synthesis and biological evaluation. Molecules. 2016;21(4):491.PubMedPubMedCentralCrossRef
15.
Salzet M, Chopin V, Baert JL, Matias I, Malecha J. Theromin, a novel leech thrombin inhibitor. J Biol Chem. 2000;275(40):30774–80.PubMedCrossRef
16.
Arocas V, Zingali RB, Guillin MC, Bon C, Jandrot-Perrus M. Bothrojaracin: a potent two-site-directed thrombin inhibitor. Biochemistry. 1996;35(28):9083–9.PubMedCrossRef
17.
Faria F, Kelen EMA, Sampaio CAM, Bon C, Duval N, Chudzinski-Tavassi AM. A new factor Xa inhibitor (lefaxin) from the Haementeria depressa leech. Thromb Haemost. 1999;82(5):1469–73.PubMedCrossRef
18.
Mao SS, Huang J, Welebob C, Neeper MP, Garsky VM, Shafer JA, et al. Identification and characterization of variants of tick anticoagulant peptide with increased inhibitory potency toward human factor Xa. Biochemistry. 1995;34(15):5098–103.PubMedCrossRef
19.
Li QQ, Yang YX, Qv JW, Hu G, Hu YJ, Xia ZN, et al. Investigation of interactions between thrombin and ten phenolic compounds by affinity capillary electrophoresis and molecular docking. J Anal Methods Chem. 2018;2018:4707609.PubMedPubMedCentral
20.
Bijak M, Ponczek MB, Nowak P. Polyphenol compounds belonging to flavonoids inhibit activity of coagulation factor X. Int J Biol Macromol. 2014;65:129–35.PubMedCrossRef
21.
Dahmer T, Berger M, Barlette AG, Reck J, Segalin J, Verza S, et al. Antithrombotic effect of chikusetsusaponin IVa isolated from Ilex paraguariensis (Maté). J Med Food. 2012;15(12):1073–80.PubMedCrossRef
22.
Ku SK, Bae JS. Antithrombotic activities of sulforaphane via inhibiting platelet aggregation and FIIa/FXa. Arch Pharm Res. 2014;37(11):1454–63.PubMedCrossRef
23.
Zhang Q, Yang YX, Li SY, Wang YL, Yang FQ, Chen H, et al. An ultrafiltration and high performance liquid chromatography coupled with diode array detector and mass spectrometry approach for screening and characterizing thrombin inhibitors from Rhizoma Chuanxiong. J Chromatogr B. 2017;1061:421–9.CrossRef
24.
Song HP, Wu SQ, Hao HP, Chen J, Lu J, Xu XJ, et al. A chemical family-based strategy for uncovering hidden bioactive molecules and multicomponent interactions in herbal medicines. Sci Rep. 2016;6:23840.PubMedPubMedCentralCrossRef
25.
Li ZY, Du Y, Yuan YL, Zhang XJ, Wang ZT, Tian X. Integrated quality evaluation strategy for multi-species resourced herb medicine of Qinjiao by metabolomics analysis and genetic comparation. Chin Med. 2020;15:16.PubMedPubMedCentralCrossRef
26.
Kellogg JJ, Todd DA, Egan JM, Raja HA, Oberlies NH, Kvalheim OM, et al. Biochemometrics for natural products research: comparison of data analysis approaches and application to identification of bioactive compounds. J Nat Prod. 2016;79(2):376–86.PubMedPubMedCentralCrossRef
27.
Yan Y, Du CH, Li ZY, Zhang M, Li J, Jia JP, et al. Comparing the antidiabetic effects and chemical profiles of raw and fermented Chinese Ge-Gen-Qin-Lian decoction by integrating untargeted metabolomics and targeted analysis. Chin Med. 2018;13:54.PubMedPubMedCentralCrossRef
28.
Zhang Q, Tan CN, Cai L, Xia FB, Gao D, Yang FQ, et al. Characterization of active antiplatelet chemical compositions of edible Citrus limon through ultra-performance liquid chromatography single quadrupole mass spectrometry-based chemometrics. Food Funct. 2018;9(5):2762–73.PubMedCrossRef
29.
Ma CH, Dastmalchi K, Flores G, Wu SB, Pedraza-Peñalosa P, Long CL, et al. Antioxidant and metabolite profiling of North American and neotropical blueberries using LC–TOF–MS and multivariate analyses. J Agr Food Chem. 2013;61(14):3548–59.CrossRef
30.
31.
Li F, Gonzalez FJ, Ma XC. LC-MS-based metabolomics in profiling of drug metabolism and bioactivation. Acta Pharm Sin B. 2012;2(2):118–25.CrossRef
32.
Rosenfeld RJ, Goodsell DS, Musah RA, Morris GM, Goodin DB, Olson AJ. Automated docking of ligands to an artificial active site: augmenting crystallographic analysis with computer modeling. J Comput Aided Mol Des. 2003;17(8):525–36.PubMedCrossRef
33.
Banner DW, Hadváry P. Crystallographic analysis at 3.0-Å resolution of the binding to human thrombin of four active site-directed inhibitors. J Biol Chem. 1991;266(30):20085–93.PubMed
34.
Roehrig S, Straub A, Pohlmann J, Lampe T, Pernerstorfer J, Schlemmer KH, et al. Discovery of the novel antithrombotic agent 5-Chloro-N-({(5S)-2-oxo-3- [4-(3-oxomorpholin-4-yl)phenyl]-1,3-oxazolidin-5-yl}methyl)thiophene-2-carboxamide (BAY 59-7939): an oral, direct factor Xa inhibitor. J Med Chem. 2005;48(19):5900–8.PubMedCrossRef
35.
Eriksson L, Jaworska J, Worth AP, Cronin MTD, McDowell RM, Gramatica P. Methods for reliability and uncertainty assessment and for applicability evaluations of classification- and regression-based QSARs. Environ Health Perspect. 2003;111(10):1361–75.PubMedPubMedCentralCrossRef
36.
Wiklund S, Johansson E, Sjöström L, Mellerowicz EJ, Edlund U, Shockcor JP, et al. Visualization of GC/TOF–MS-based metabolomics data for identification of biochemically interesting compounds using OPLS class models. Anal Chem. 2008;80(1):115–22.PubMedCrossRef
37.
Zhu ZY, Zhang H, Zhao L, Dong X, Li X, Chai YF, et al. Rapid separation and identification of phenolic and diterpenoid constituents from Radix Salvia miltiorrhizae by high-performance liquid chromatography diode-array detection, electrospray ionization time-of-flight mass spectrometry and electrospray ionization quadrupole ion trap mass spectrometry. Rapid Commun Mass Spectrom. 2007;21(12):1855–65.PubMedCrossRef
38.
Liang WY, Chen WJ, Wu LF, Li S, Qi Q, Cui YP, et al. Quality evaluation and chemical markers screening of salvia miltiorrhiza Bge. (Danshen) based on HPLC fingerprints and HPLC-MSn coupled with chemometrics. Molecules. 2017;22(3):478.PubMedCentralCrossRef
39.
Yang M, Liu AH, Guan SH, Sun JH, Xu M, Guo DA. Characterization of tanshinones in the roots of Salvia miltiorrhiza (Dan-shen) by high-performance liquid chromatography with electrospray ionization tandem mass spectrometry. Rapid Commun Mass Spectrom. 2006;20(8):1266–80.PubMedCrossRef
40.
Zhang L, Chen HX, Chen Y. EIS-MS Performance and fingerprint of lipophilic constituents from Radix Salviae Miltiorrhiza. Chin Herb Med. 2006;01:122–5.
41.
Qu C, Pu ZJ, Zhou GS, Wang J, Zhu ZH, Yue SJ, et al. Comparative analysis of main bio-active components in the herb pair Danshen-Honghua and its single herbs by ultra high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry. J Sep Sci. 2017;40(17):3392–401.PubMedCrossRef
42.
Böhm M, Stürzebecher J, Klebe G. Three-dimensional quantitative structure–activity relationship analyses using comparative molecular field analysis and comparative molecular similarity indices analysis to elucidate selectivity differences of inhibitors binding to Trypsin, Thrombin, and Factor Xa. J Med Chem. 1999;42:458–77.PubMedCrossRef
43.
Nar H. The role of structural information in the discovery of direct thrombin and factor Xa inhibitors. Trends Pharmacol Sci. 2012;33(5):279–88.PubMedCrossRef
44.
Vaz RJ, McLean LR, Pelton JT. Evaluation of proposed modes of binding of (2S)-2-[4-[[(3S)-1-acetimidoyl3-pyrrolidinyl]oxy]phenyl]-3-(7-amidino-2-naphthyl)propanoic acid hydrochloride and some analogs to Factor Xa using a comparative molecular field analysis. J Comp Aided Mol. 1998;12(2):99–110.CrossRef
45.
Lu J, Song HP, Li P, Zhou P, Dong X, Chen J. Screening of direct thrombin inhibitors from Radix Salviae Miltiorrhizae by a peak fractionation approach. J Pharm Biomed Anal. 2015;109:85–90.PubMedCrossRef
Metadata
Title
LC–MS-based multivariate statistical analysis for the screening of potential thrombin/factor Xa inhibitors from Radix Salvia Miltiorrhiza
Authors
Yi-Yao Yang
Zhao-Yu Wu
Hao Zhang
Shi-Jun Yin
Fang-Bo Xia
Qian Zhang
Jian-Bo Wan
Jian-Li Gao
Feng-Qing Yang
Publication date
01-12-2020
Publisher
BioMed Central
Published in
Chinese Medicine / Issue 1/2020
Electronic ISSN: 1749-8546
DOI
https://doi.org/10.1186/s13020-020-00320-2

Other articles of this Issue 1/2020

Chinese Medicine 1/2020 Go to the issue

Research

Integrated assessment of medicinal rhubarb by combination of delayed luminescence and HPLC fingerprint with emphasized on bioactivities based quality control

Research

Effect of Loureirin B on Crohn’s disease rat model induced by TNBS via IL-6/STAT3/NF-κB signaling pathway

Research

Anti-dengue virus serotype 2 activity of tannins from porcupine dates

Research

The curative effects of the traditional Chinese herbal medicine “Jinchuang ointment” on excisional wounds

Review

The involvement of human organic anion transporting polypeptides (OATPs) in drug-herb/food interactions

Research

A comparative study of aged and contemporary Chinese herbal materials by using delayed luminescence technique