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1Traditional Medicine Research  2020, Vol. 5 Issue (3): 145-159    DOI: 10.12032/TMR20200324170
Special Issue on Infectious Diseases and Public Health     
Network pharmacology studies on the effect of Chai-Ling decoction in coronavirus disease 2019
Lu Yang1#, Yu-Ting Li2#, Jing Miao3, Li Wang4, Hui Fu5, Qin Li6, Wei-Bo Wen6, Zhai-Yi Zhang5, Rui-Wen Song7, Xiang-Guo Liu8, Hong-Wu Wang2,*(), Huan-Tian Cui8,*()
1Graduate School, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
2College of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
3Department of Integrated Traditional and Western Medicine, Tianjin Second People’s Hospital, Tianjin 300192, China;
4Department of Pharmacy, Tianjin Second People’s Hospital, Tianjin 300192, China;
5College of Integrated Chinese and Western Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
6Department of Endocrinology in Yunnan Provincial Hospital of Traditional Chinese Medicine, Kunming 650021, China
7School of Management, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
8Shandong Provincial Key Laboratory of Animal Cell and Developmental Biology, School of Life Sciences, Shandong University, Qingdao 250100, China.
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The current study applied network pharmacology analysis and molecular docking method to study the potential mechanisms of Chai-Ling decoction (CLD), an empirical formula derived from the classic ancient prescription Xiao-Chai-Hu (XCH) decoction and Wu-Ling-San (WLS), on coronavirus disease 2019 (COVID-19).


The classic ancient prescription XCH and WLS decoctions originated from the ancient book of Chinese medicine Shang Han Za Bing Lun (Treatise on Cold Damage Disorders, 200-210 C.E.), written by Zhang Zhongjing. Previous studies have demonstrated that XCH can alleviate fever, cough, and fatigue, which were the primary clinical outcomes of COVID-19. Besides, WLS decoction has shown apparent effects on attenuating gastrointestinal symptoms. CLD, derived from a modification of XCH and WLS decoctions, is used to treat the early-stage of COVID-19 in the Prevention and Treatment Guidelines of Damp-Heat Syndrome of “Taiyin” Lung (respiratory system in the theory of traditional Chinese medicine) Epidemic Disease (coronavirus pneumonia). However, the mechanisms of action of CLD in COVID-19 remain unclear.


Background: Chai-Ling decoction (CLD), derived from a modification of Xiao-Chai-Hu (XCH) decoction and Wu-Ling-San (WLS) decoction, has been used to treat the early-stage of coronavirus disease 2019 (COVID-19). However, the mechanisms of CLD in COVID-19 remain unknown. In this study, the potential mechanisms of CLD in COVID-19 were preliminarily investigated based on network pharmacology and molecular docking method. Methods: Initially, the active components and targets of CLD were screened based on Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and PharmMapper database. The targets of COVID-19 were obtained from GeneCards database. The protein-protein interaction network was established using STRING database to analyze the key targets. Gene Oncology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes analysis were also conducted to evaluate the pathways related to the targets of CLD on COVID-19. Moreover, the compound-target-pathway network was established using Cytoscape 3.2.7. Subsequently, the molecular docking method was performed to select the active compounds with high binding affinity on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and angiotensin-converting enzyme 2 (ACE2), which is the key target of SARS-CoV-2 in entering target cells. The possible binding sites were also visualized by a three-dimensional graph. Results: Network pharmacology analysis showed that there were 106 active components and 160 targets of CLD. Additionally, 251 targets related to COVID-19 were identified, and 24 candidates of CLD on COVID-19 were selected. A total of 283 GO terms of CLD on COVID-19 were identified, and 181 pathways were screened based on GO and Kyoto Encyclopedia of Genes and Genomes analyses. CLD might alleviate the inflammatory response and improve lung injury to treat COVID-19 through interleukin 17 signaling, T helper cell 17 differentiation, tumor necrosis factor signaling, and hypoxia inducible factor-1 signaling. Besides, molecular docking indicated that beta-sitosterol, kaempferol, and stigmasterol were the top three candidates in CLD with the highest affinity to SARS-CoV-2 and ACE2. Conclusion: Our study identifies the potential mechanisms of CLD on COVID-19 and beta-sitosterol, kaempferol, and stigmasterol may be the key compounds that exert antiviral effects against SARS-CoV-2.

Key wordsChai-Ling decoction      Coronavirus disease 2019      Network pharmacology      Molecular docking      Severe acute respiratory syndrome coronavirus 2      Angiotensin-converting enzyme 2     
Published: 13 April 2020
Fund:  This study was supported by university-level project on curing disease in 2018 of Tianjin University of Traditional Chinese Medicine (XJ201801).
Corresponding Authors: Hong-Wu Wang, Huan-Tian Cui   
About author: #These authors are co-first authors on this work.
Cite this article:

Lu Yang, Yu-Ting Li, Jing Miao, Li Wang, Hui Fu, Qin Li, Wei-Bo Wen, Zhai-Yi Zhang, Rui-Wen Song, Xiang-Guo Liu, Hong-Wu Wang, Huan-Tian Cui. Network pharmacology studies on the effect of Chai-Ling decoction in coronavirus disease 2019. 1Traditional Medicine Research, 2020, 5(3): 145-159. doi: 10.12032/TMR20200324170


MOL_ID Molecule_name OB (%) DL Herb Node
MOL000006 Luteolin 36.16 0.25 FF M1
MOL000098 Quercetin 46.43 0.28 FF, BR, GRER M2
MOL000173 Wogonin 30.68 0.23 FF, SR M3
MOL000358 Beta-sitosterol 36.91 0.75 FF, CR, SR M4
MOL000422 Kaempferol 41.88 0.24 FF, BR, GRER M5
MOL003295 (+)-Pinoresinol monomethyl ether 53.08 0.57 FF M6
MOL003305 Phillyrin 36.4 0.86 FF M7
MOL003306 ACon1_001697 85.12 0.57 FF M8
MOL003322 Forsythinol 81.25 0.57 FF M9
MOL003330 (-)-Phillygenin 95.04 0.57 FF M10
MOL003347 Hyperforin 44.03 0.6 FF M11
MOL003348 Adhyperforin 44.03 0.61 FF M12
MOL000359 Sitosterol 36.91 0.75 SD, AR, CR, SR, GRER M13
MOL000830 Alisol B 34.47 0.82 AR M14
MOL000831 Alisol B monoacetate 35.58 0.81 AR M15
MOL000832 Alisol B 23-acetate 32.52 0.82 AR M16
MOL000853 Alisol B 36.76 0.82 AR M17
MOL000854 Alisol C 32.7 0.82 AR M18
MOL000856 Alisol C monoacetate 33.06 0.83 AR M19
MOL000816 Ergosta-7,22-dien-3-one 44.88 0.72 PO M20
MOL000817 Ergosta-5,7,22-trien-3-ol 46.18 0.72 PO M21
MOL000820 Polyporusterone E 45.71 0.85 PO M22
MOL000822 Polyporusterone G 33.43 0.81 PO M23
MOL011169 Peroxyergosterol 44.39 0.82 PO M24
MOL000276 7,9(11)-Dehydropachymic acid 35.11 0.81 P M25
MOL000283 Ergosterol peroxide 40.36 0.81 P M26
MOL000287 3Beta-hydroxy-24-methylene-8-lanostene-21-oic acid 38.7 0.81 P M27
MOL000289 Pachymic acid 33.63 0.81 P M28
MOL000290 Poricoic acid A 30.61 0.76 P M29
MOL000291 Poricoic acid B 30.52 0.75 P M30
MOL000292 Poricoic acid C 38.15 0.75 P M31
MOL000296 Hederagenin 36.91 0.75 P M32
MOL000300 Dehydroeburicoic acid 44.17 0.83 P M33
MOL000021 14-Acetyl-12-senecioyl-2E,8E,10E-atractylentriol 60.31 0.31 AMR M34
MOL000022 14-Acetyl-12-senecioyl-2E,8Z,10E-atractylentriol 63.37 0.3 AMR M35
MOL000049 3β-Acetoxyatractylone 54.07 0.22 AMR M36
MOL000072 8β-Ethoxy atractylenolide Ⅲ 35.95 0.21 AMR M37
MOL000073 ent-Epicatechin 48.96 0.24 CR M38
MOL000492 (+)-Catechin 54.83 0.24 CR M39
MOL001736 (-)-Taxifolin 60.51 0.27 CR M40
MOL_ID Molecule_name OB (%) DL Herb Node
MOL000228 (2R)-7-Hydroxy-5-methoxy-2-
55.23 0.2 SR M41
MOL000525 Norwogonin 39.4 0.21 SR M42
MOL000552 5,2'-Dihydroxy-6,7,8-trimethoxyflavone 31.71 0.35 SR M43
MOL001689 Acacetin 34.97 0.24 SR M44
MOL002714 Baicalein 33.52 0.21 SR M45
MOL002908 5,8,2'-Trihydroxy-7-methoxyflavone 37.01 0.27 SR M46
MOL002909 5,7,2,5-Tetrahydroxy-8,6-
33.82 0.45 SR M47
MOL002917 5,2',6'-Trihydroxy-7,8-dimethoxyflavone 45.05 0.33 SR M48
MOL002925 5,7,2',6'-Tetrahydroxyflavone 37.01 0.24 SR M49
MOL002927 Skullcapflavone II 69.51 0.44 SR M50
MOL002928 Oroxylin a 41.37 0.23 SR M51
MOL002932 Panicolin 76.26 0.29 SR M52
MOL002933 5,7,4'-Trihydroxy-8-methoxyflavone 36.56 0.27 SR M53
MOL002934 Neobaicalein 104.3 0.44 SR M54
MOL000354 Isorhamnetin 49.6 0.31 BR, GRER M55
MOL000449 Stigmasterol 43.83 0.76 BR M56
MOL002776 Baicalin 40.12 0.75 BR M57
MOL004598 3,5,6,7-Tetramethoxy-2-(3,4,5-
31.97 0.59 BR M58
MOL004609 Areapillin 48.96 0.41 BR M59
MOL004648 Troxerutin 31.6 0.28 BR M60
MOL000239 Jaranol 50.83 0.29 GRER M61
MOL000392 Formononetin 69.67 0.21 GRER M62
MOL000417 Calycosin 47.75 0.24 GRER M63
MOL000497 Licochalcone a 40.79 0.29 GRER M64
MOL000500 Vestitol 74.66 0.21 GRER M65
MOL001484 Inermine 75.18 0.54 GRER M66
MOL001792 DFV 32.76 0.18 GRER M67
MOL002311 Glycyrol 90.78 0.67 GRER M68
MOL002565 Medicarpin 49.22 0.34 GRER M69
MOL003896 7-Methoxy-2-methyl isoflavone 42.56 0.2 GRER M70
MOL004328 Naringenin 59.29 0.21 GRER M71
MOL004808 Glyasperin B 65.22 0.44 GRER M72
MOL004810 Glyasperin F 75.84 0.54 GRER M73
MOL004811 Glyasperin C 45.56 0.4 GRER M74
MOL004815 (E)-1-(2, 4-Dihydroxyphenyl)-3-(2, 2-dimethylchromen-6-yl)prop-2-en-1-one 39.62 0.35 GRER M75
MOL004827 Semilicoisoflavone B 48.78 0.55 GRER M76
MOL004828 Glepidotin A 44.72 0.35 GRER M77
MOL004829 Glepidotin B 64.46 0.34 GRER M78
MOL004833 Phaseolinisoflavan 32.01 0.45 GRER M79
MOL004841 Licochalcone B 76.76 0.19 GRER M80
MOL_ID Molecule_name OB (%) DL Herb Node
MOL004848 Licochalcone G 49.25 0.32 GRER M81
MOL004855 Licoricone 63.58 0.47 GRER M82
MOL004856 Gancaonin A 51.08 0.4 GRER M83
MOL004879 Glycyrin 52.61 0.47 GRER M84
MOL004883 Licoisoflavone 41.61 0.42 GRER M85
MOL004884 Licoisoflavone B 38.93 0.55 GRER M86
MOL004885 Licoisoflavanone 52.47 0.54 GRER M87
MOL004903 Liquiritin 65.69 0.74 GRER M88
MOL004908 Glabridin 53.25 0.47 GRER M89
MOL004910 Glabranin 52.9 0.31 GRER M90
MOL004911 Glabrene 46.27 0.44 GRER M91
MOL004912 Glabrone 52.51 0.5 GRER M92
MOL004915 Eurycarpin A 43.28 0.37 GRER M93
MOL004917 Glycyroside 37.25 0.79 GRER M94
MOL004924 (-)-Medicocarpin 40.99 0.95 GRER M95
MOL004949 Isolicoflavonol 45.17 0.42 GRER M96
MOL004957 HMO 38.37 0.21 GRER M97
MOL004959 1-Methoxyphaseollidin 69.98 0.64 GRER M98
MOL004961 Quercetin der. 46.45 0.33 GRER M99
MOL005000 Gancaonin G 60.44 0.39 GRER M100
MOL005001 Gancaonin H 50.1 0.78 GRER M101
MOL005007 Glyasperins M 72.67 0.59 GRER M102
MOL005008 Glycyrrhiza flavonol A 41.28 0.6 GRER M103
MOL005012 Licoagroisoflavone 57.28 0.49 GRER M104
MOL005013 18α-Hydroxyglycyrrhetic acid 41.16 0.71 GRER M105
MOL005020 Dehydroglyasperins C 53.82 0.37 GRER M106
Table 1 Information of active compounds in CLD
Figure 1 Common targets of CLD in COVID-19. CLD, Chai-Ling decoction; COVID-19, coronavirus disease 2019.
Figure 2 PPI network of target proteins. PPI, protein-protein interaction.
Figure 3 GO enrichment analysis of the common targets. GO, Gene oncology.
No. Pathways Numbers of genes - log (p)
1 Human cytomegalovirus infection 12 22.22
2 IL-17 signaling pathway 10 21.29
3 Tuberculosis 10 18.58
4 Endocrine resistance 9 18.54
5 C-type lectin receptor signaling pathway 9 18.32
6 HIF-1 signaling pathway 9 18.14
7 TNF signaling pathway 9 18.04
8 NOD-like receptor signaling pathway 9 16.30
9 T cell receptor signaling pathway 8 15.90
10 Th17 cell differentiation 8 15.78
Table 2 KEGG enrichment analysis of the common targets
Figure 4 Compound-target-pathway network of CLD in COVID-19. Red nodes represent the genes, orange nodes represent pathways, light green nodes represent the active ingredients of Chaihu (Bupleuri Radix), purple nodes represent active ingredient of Huangqin (Scutellariae radix), light yellow nodes represent active ingredients of Lianqiao (Forsythiae Fructus), light blue nodes represent active ingredients of Gancao (Glycyrrhizae Radix et Rhizoma), and pink nodes represent the active ingredients of Guizhi (Cinnamomi Ramulus), Baizhu (Atractylodis Macrocephalae Rhizoma), Fuling (Poria), Zhuling (Polyporus), Zexie (Alismatis Rhizoma), Shishangbai (Selaginella Doederleinii).
CLD, Chai-Ling decoction; COVID-19, coronavirus disease 2019.
Compound Molecular formula Binding energy values (SARS-CoV-2) Binding energy values (ACE2)
Luteolin C15H10O6 - 7.4 - 8.8
Quercetin C15H10O7 - 7.5 - 9.0
Wogonin C16H12O5 - 6.7 - 8.8
Mairin C30H48O3 - 7.4 - 9.6
Beta-sitosterol C30H52O - 8.1 - 10.9
Kaempferol C15H10O6 - 7.8 - 10.4
Arctiin C27H34O11 - 7.3 - 9.9
Bicuculline C20H17NO6 - 7.5 - 8.6
(3R,4R)-3,4-bis[(3,4-dimethoxyphenyl)methyl]oxolan-2-one C13H22O - 6.5 - 8.7
(+)-Pinoresinol monomethyl ether C27H34O11 - 6.9 - 8.9
Phyllyrin C27H34O11 - 7.1 - 9.4
Forsythinol C21H24O6 - 4.8 - 5.6
(-)-Phillygenin C21H24O6 - 6.9 - 9.0
β-Amyrin acetate C32H52O2 - 7.7 - 6.5
Hyperforin C35H52O4 - 5.9 - 9.1
Onjixanthone I C16H14O6 - 6.5 - 7.9
Stigmasterol C29H48O - 7.7 - 9.8
Yangambin C24H30O8 - 6.3 - 8.2
Alisol b C28H44O4 - 7.6 - 9.7
Alisol, b, 23-acetate C31H47O6 - 6.9 - 9.7
Arbidol C22H26BrClN2O3S - 6.5 - 8.2
Atazanavir C38H52N6O7 - 7.7 - 12.4
Darunavir C27H37N3O7S - 7.6 - 9.6
Indinavir C36H47N5O4 - 8.2 - 10.9
Kyprolis C40H57N5O7 - 7.3 - 10.2
Lopinavir C37H48N4O5 - 9.1 - 10.8
Remdesivir C27H35N6O8P - 7.5 - 11.0
Ritonavir C37H48N6O5S2 - 8.0 - 9.9
Saquinavir C38H50N6O5 - 8.9 - 11.7
Tipranavir C31H33F3N2O5S - 7.7 - 11.7
Table 4 Molecular docking table of ACE2 and SARS-CoV-2 with 20 active components in CLD and 10 Western medicine compounds
Figure 5 Molecular docking diagram of ACE2 with β-sitosterol, kaempferol, and stigmasterol. ACE2, angiotensin-converting enzyme 2.
Figure 6 Molecular docking diagram of SARS-CoV-2 with β-sitosterol, kaempferol, and stigmasterol. SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.
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