Three salvianolic acids inhibit 2019‐nCoV spike pseudovirus viropexis by binding to both its RBD and receptor ACE2

<jats:title>Abstract</jats:title><jats:p>Since December 2019, the new coronavirus (also known as severe acute respiratory syndrome coronavirus 2 [SARS‐CoV‐2, 2019‐nCoV])—induced disease, COVID‐19, has spread rapidly worldwide. Studies have reported that the traditional Chinese medicine <jats:italic>Salvia miltiorrhiza</jats:italic> possesses remarkable antiviral properties; however, the anti‐coronaviral activity of its main components, salvianolic acid A (SAA), salvianolic acid B (SAB), and salvianolic acid C (SAC) is still debated. In this study, we used Cell Counting Kit‐8 staining and flow cytometry to evaluate the toxicity of SAA, SAB, and SAC on ACE2 (angiotensin‐converting enzyme 2) high‐expressing HEK293T cells (ACE2<jats:sup>h</jats:sup> cells). We found that SAA, SAB, and SAC had a minor effect on the viability of ACE2<jats:sup>h</jats:sup> cells at concentrations below 100 μM. We further evaluated the binding capacity of SAA, SAB, and SAC to ACE2 and the spike protein of 2019‐nCoV using molecular docking and surface plasmon resonance. They could bind to the receptor‐binding domain (RBD) of the 2019‐nCoV with a binding constant (<jats:italic>K</jats:italic><jats:sub>D</jats:sub>) of (3.82 ± 0.43) e−6 M, (5.15 ± 0.64)e−7 M, and (2.19 ± 0.14)e‐6 M; and bind to ACE2 with <jats:italic>K</jats:italic><jats:sub>D</jats:sub> (4.08 ± 0.61)e−7 M, (2.95 ± 0.78)e−7 M, and (7.32 ± 0.42)e−7 M, respectively. As a result, SAA, SAB, and SAC were determined to inhibit the entry of 2019‐nCoV Spike pseudovirus with an EC<jats:sub>50</jats:sub> of 11.31, 6.22, and 10.14 μM on ACE2<jats:sup>h</jats:sup> cells, respectively. In conclusion, our study revealed that three Salvianolic acids can inhibit the entry of 2019‐nCoV spike pseudovirus into ACE2<jats:sup>h</jats:sup> cells by binding to the RBD of the 2019‐nCoV spike protein and ACE2 protein.</jats:p>