Blocking Effect of Demethylzeylasteral on the Interaction between Human ACE2 Protein and SARS-CoV-2 RBD Protein Discovered Using SPR Technology.

Zhi-Ling Zhu,Yi-Tong Liu,Xiao-Dan Qiu,Ting Zhao,Zhuo-Rong Li,Shuo Wu,Zhong-Hao Sun,Guang-Zhi Shan

doi:10.3390/molecules26010057

Abstract

The novel coronavirus disease (2019-nCoV) has been affecting global health since the end of 2019, and there is no sign that the epidemic is abating. Targeting the interaction between the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein and the human angiotensin-converting enzyme 2 (ACE2) receptor is a promising therapeutic strategy. In this study, surface plasmon resonance (SPR) was used as the primary method to screen a library of 960 compounds. A compound 02B05 (demethylzeylasteral, CAS number: 107316-88-1) that had high affinities for S-RBD and ACE2 was discovered, and binding affinities (KD, μM) of 02B05-ACE2 and 02B05-S-RBD were 1.736 and 1.039 μM, respectively. The results of a competition experiment showed that 02B05 could effectively block the binding of S-RBD to ACE2 protein. Furthermore, pseudovirus infection assay revealed that 02B05 could inhibit entry of SARS-CoV-2 pseudovirus into 293T cells to a certain extent at nontoxic concentration. The compoundobtained in this study serve as references for the design of drugs which have potential in the treatment of COVID-19 and can thus accelerate the process of developing effective drugs to treat SARS-CoV-2 infections.

Highlights

Received: 17 November 2020 Accepted: 22 December 2020 Published: 24 December 2020Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global outbreak of coronavirus disease 2019 (COVID-19) [1,2]
The angiotensinconverting enzyme 2 (ACE2)-His protein was first captured as a ligand and S-receptor binding domain (RBD)-mFc was used as an analyte to evaluate the activity of the two proteins
The measured S-RBD maximum response signal was 94.2 response units (RUs), which did not differ significantly from the theoretical Rmax of 87.7 RU. These results demonstrated that the activity of both the proteins remained good in the running buffer

Summary

Introduction

Received: 17 November 2020 Accepted: 22 December 2020 Published: 24 December 2020Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global outbreak of coronavirus disease 2019 (COVID-19) [1,2]. Received: 17 November 2020 Accepted: 22 December 2020 Published: 24 December 2020. Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global outbreak of coronavirus disease 2019 (COVID-19) [1,2]. SARS-CoV-2 infection can cause symptoms such as fever, dry cough and dyspnea. Unlike SARS, upper respiratory symptoms and intestinal presentations of COVID-19 are not common [1]. The S protein of SARS-CoV-2 mainly mediates binding with the membrane receptor on the host cells (angiotensin-converting enzyme 2; ACE2) and facilitates viral entry into host cells [5,6,7,8,9,10]. Amino acid variations in this region cause changes in the species’ tropism and infection characteristics [11,12]

Methods

Results

Conclusion