title: Identification of berberine as a direct thrombin inhibitor from traditional Chinese medicine through structural, functional and binding studies.
abstract: Thrombin acts as a key enzyme in the blood coagulation cascade and represents a potential drug target for the treatment of several cardiovascular diseases. The aim of this study was to identify small-molecule direct thrombin inhibitors from herbs used in traditional Chinese medicine (TCM). A pharmacophore model and molecular docking were utilized to virtually screen a library of chemicals contained in compositions of traditional Chinese herbs, and these analyses were followed by in vitro bioassay validation and binding studies. Berberine (BBR) was first confirmed as a thrombin inhibitor using an enzymatic assay. The BBR IC50value for thrombin inhibition was 2.92 μM. Direct binding studies using surface plasmon resonance demonstrated that BBR directly interacted with thrombin with a KDvalue of 16.39 μM. Competitive binding assay indicated that BBR could bind to the same argartroban/thrombin interaction site. A platelet aggregation assay demonstrated that BBR had the ability to inhibit thrombin-induced platelet aggregation in washed platelets samples. This study proved that BBR is a direct thrombin inhibitor that has activity in inhibiting thrombin-induced platelet aggregation. BBR may be a potential candidate for the development of safe and effective thrombin-inhibiting drugs.
authors.affiliation: Beijing Key Lab of Traditional Chinese Medicine (TCM) Collateral Disease Theory Research, School of Traditional Chinese Medicine, Capital Medical University, 10 Youanmen, Xitoutiao, Beijing 100069, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of TCM-Information Engineer of State Administration of TCM, School of Chinese Materia Medica, Beijing University of Chinese Medicine, 6 Central Ring South Road, Wangjing, Beijing 100102, China.
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Core Facilities Center, Capital Medical University, 10 Youanmen, Xitoutiao, Beijing 100069, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Beijing Key Lab of Traditional Chinese Medicine (TCM) Collateral Disease Theory Research, School of Traditional Chinese Medicine, Capital Medical University, 10 Youanmen, Xitoutiao, Beijing 100069, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Wu
authors.affiliation: Beijing Key Lab of Traditional Chinese Medicine (TCM) Collateral Disease Theory Research, School of Traditional Chinese Medicine, Capital Medical University, 10 Youanmen, Xitoutiao, Beijing 100069, China.
authors.forename: Hantian
authors.initials: H
authors.lastname: Fan
authors.affiliation: Key Laboratory of TCM-Information Engineer of State Administration of TCM, School of Chinese Materia Medica, Beijing University of Chinese Medicine, 6 Central Ring South Road, Wangjing, Beijing 100102, China.
authors.forename: Yanjiang
authors.initials: Y
authors.lastname: Qiao
mesh_terms: D001599:Berberine / Q000737:chemistry* / Q000494:pharmacology; D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism; D006801:Humans; D008516:Medicine, Chinese Traditional; D062105:Molecular Docking Simulation; D010974:Platelet Aggregation / Q000187:drug effects; D013917:Thrombin / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001599:Berberine; D013917:Thrombin