2. Academic Validation
  3. Biflavones from Ginkgo biloba as inhibitors of human thrombin

Biflavones from Ginkgo biloba as inhibitors of human thrombin

  • Bioorg Chem. 2019 Nov;92:103199. doi: 10.1016/j.bioorg.2019.103199.
Tian-Ran Chen 1,  Ling-Hua Wei 2,  Xiao-Qing Guan 3,  Chao Huang 3,  Zhe-Yi Liu 4,  Fang-Jun Wang 4,  Jie Hou 5,  Qiang Jin 3,  Yi-Fan Liu 6,  Pei-Hao Wen 6,  Shui-Jun Zhang 6,  Guang-Bo Ge 7,  Wen-Zhi Guo 8
Affiliations

Affiliations

  • 1 Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, China; Henan Key Laboratory of Digestive Organ Transplantation, China; Zhengzhou Key Laboratory of Hepatobiliary & Pancreatic Diseases and Organ Transplantation, China; Open and Key Laboratory of Hepatobiliary & Pancreatic Surgery and Digestive Organ Transplantation at Henan Universities, Zhengzhou 450001, China; Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
  • 2 Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Taixing People's Hospital, 1 Changzheng Road, Taixing, Jiangsu 225400, China.
  • 3 Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
  • 4 CAS Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
  • 5 College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
  • 6 Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, China; Henan Key Laboratory of Digestive Organ Transplantation, China; Zhengzhou Key Laboratory of Hepatobiliary & Pancreatic Diseases and Organ Transplantation, China; Open and Key Laboratory of Hepatobiliary & Pancreatic Surgery and Digestive Organ Transplantation at Henan Universities, Zhengzhou 450001, China.
  • 7 Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. Electronic address: [email protected].
  • 8 Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, China; Henan Key Laboratory of Digestive Organ Transplantation, China; Zhengzhou Key Laboratory of Hepatobiliary & Pancreatic Diseases and Organ Transplantation, China; Open and Key Laboratory of Hepatobiliary & Pancreatic Surgery and Digestive Organ Transplantation at Henan Universities, Zhengzhou 450001, China. Electronic address: [email protected].
PMID: 31446241 DOI: 10.1016/j.bioorg.2019.103199
Abstract

Ginkgo Biloba leaf extract has been widely used for the prevention and treatment of thrombosis and Cardiovascular Disease in both eastern and western countries, but the bioactive constituents and the underlying mechanism of anti-thrombosis have not been fully characterized. The purpose of this study was to investigate the inhibitory effects of major constituents in Ginkgo biloba on human Thrombin, a key serine protease regulating the blood coagulation cascade and the processes of thrombosis. To this end, a fluorescence-based biochemical assay was used to assay the inhibitory effects of sixteen major constituents from Ginkgo biloba on human Thrombin. Among all tested natural compounds, four Biflavones (ginkgetin, isoginkgetin, bilobetin and amentoflavone), and five Flavonoids (luteolin, apigenin, quercetin, kaempferol and isorhamnetin) were found with Thrombin inhibition activity, with the IC50 values ranging from 8.05 μM to 82.08 μM. Inhibition kinetic analyses demonstrated that four Biflavones were mixed inhibitors against thrombin-mediated Z-GGRAMC acetate hydrolysis, with the Ki values ranging from 4.12 μM to 11.01 μM. Molecular docking method showed that the four Biflavones could occupy the active cavity with strong interactions of salt bridges and hydrogen bonds. In addition, mass spectrometry-based lysine labeling reactivity assay suggested that the Biflavones could bind on human Thrombin at exosite I rather than exosite II. All these findings suggested that the Biflavones in Ginkgo biloba were naturally occurring inhibitors of human Thrombin, and these compounds could be used as lead compounds for the development of novel Thrombin inhibitors with improved efficacy and high safety profiles.

Keywords

Biflavones; Ginkgo biloba; Inhibitory effects; Proteolytic activity; Thrombin.

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