key: cord-0716308-bk6imjl2
authors: Hou, Yajing; Ge, Shuai; Li, Xiaowei; Wang, Cheng; He, Huaizhen; He, Langchong
title: Testing of the inhibitory effects of loratadine and desloratadine on SARS-CoV-2 spike pseudotyped virus viropexis
date: 2021-02-18
journal: Chem Biol Interact
DOI: 10.1016/j.cbi.2021.109420
sha: 00945745d3068c07c6a4ad340370feb8438cba9e
doc_id: 716308
cord_uid: bk6imjl2

Currently, there is an urgent need to find a treatment for the highly infectious coronavirus disease (COVID-19). However, the development of a new, effective, and safe vaccine or drug often requires years and poses great risks. At this critical stage, there is an advantage in using existing clinically approved drugs to treat COVID-19. In this study, in vitro severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) spike pseudotyped viral infection experiments indicated that histamine H(1) antagonists loratadine (LOR) and desloratadine (DES) could prevent entry of the pseudotyped virus into ACE2-overexpressing HEK293T cells and showed that DES was more effective. Further binding experiments using cell membrane chromatography and surface plasmon resonance demonstrated that both antagonists could bind to ACE2 and that the binding affinity of DES was much stronger than that of LOR. Molecular docking results elucidated that LOR and DES could bind to ACE2 on the interface of the SARS-CoV-2-binding area. Additionally, DES could form one hydrogen bond with LYS31 but LOR binding relied on non-hydrogen bonds. To our knowledge, this study is the first to demonstrate the inhibitory effect of LOR and DES on SARS-CoV-2 spike pseudotyped virus viropexis by blocking spike protein–ACE2 interaction. This study may provide a new strategy for finding an effective therapeutic option for COVID-19.

December 2019, this coronavirus has rapidly spread to many countries worldwide. 58 Because of the rapid and global spreading of SARS-CoV-2, COVID-19 has been 59 identified as a public health emergency of international concern [3-5]. As a virus 60 belonging to the order Nidovirales and family Coronaviridae, SARS-CoV-2 requires 61 the interaction of the viral spike (S) glycoprotein with angiotensin-converting enzyme 62 2 (ACE2) to enter cells [6] [7] [8] . The viral spike protein is a trimer, and each monomer 63 contains two subunits, S1 and S2. During the infection period, S1 is responsible for 64 viral attachment to the host cell surface through a receptor-binding domain, whereas 65 S2 is required for the fusion of the viral and cellular membranes [9, 10] . Recent 66 studies have verified that binding to ACE2 is a critical initial step for the entry of 67 SARS-CoV-2 into target cells [11, 12] . Therefore, blocking or manipulating this 68 binding process may prove very effective in defeating the virus. antagonists LOR and DES are on the top of these SARS-CoV-2-inhibiting drugs [29] . 101 Therefore, in this study, we evaluated the inhibitory effects and preliminary 102 mechanism of action of LOR and DES on SARS-CoV-2 viropexis, hoping to find 103 effective agents in against SARS-CoV-2 infection. All statistical analyses of the data were carried out using GraphPad 5.0 Prism software. 168 All of the results were represented as the mean ± standard derivation (SD) from three 169 independent experiments. * represents p-value <0.05; ** represents p-value <0.01; 170 *** represents p-value <0.001 by Student's t-test. The expression of ACE2 protein in ACE2 h cells was significantly higher than that in 174 HEK293T cells, indicating a successful construction of ACE2 h cells (Fig. 1A) . The indicating that DES had much better binding to the ACE2 receptor than LOR (Fig.   224 3A). Additionally, surface plasmon resonance (SPR) method was used to detect the 225 binding affinity of LOR and DES with ACE2. The response value of DES was easily 226 observed to be higher than that of LOR at the same concentration, with binding 227 constants of (9.13 ± 0.67) × 10 -6 M and (1.02 ± 0.38) × 10 -7 M, respectively (Fig. 3B) . 228 This result revealed that the binding affinity between DES and ACE2 was much 229 stronger than that between LOR and ACE2, which is in accordance with the CMC 

To explain the different activities of LOR and DES, molecular docking was used for 238 the virtual evaluation of their binding character with ACE2 (PDB code: 6M0J). The 239 contacting residues of ACE2 and SARS-CoV-2 spike proteins were labeled at the 240 ACE2 interfaces (Fig. 4B) . Docking results showed that the position of LOR was far 241 away from the active site of ACE2 and their interaction was mainly based on 242 non-hydrogen bonds (Fig. 4C) . However, after removing the substituent group in of them can bind with ACE2 protein, but also revealed that the binding affinity 278 between DES and ACE2 was much stronger than that between LOR and ACE2. 

Asymptomatic carrier state, acute respiratory disease, and pneumonia due to 351 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2): Facts and myths

COVID-19): what we know? J

A pneumonia 359 outbreak associated with a new coronavirus of probable batorigin

A new coronavirus associated 362 with human respiratory disease in china

A novel coronavirus from patients with pneumonia in China

Comparing the Binding Interactions in the 368 usage for SARS-CoV-2 and other lineage B betacoronaviruses

CoV-2 Cell entry depends on ACE2 and TMPRSS2 and is 388 blocked by a clinically proven protease inhibitor

Angiotensin-converting enzyme 2 (ace2) receptor and sars-cov-2: potential 391 therapeutic targeting

Development of vaccines for sars-cov-2, 393 F1000 Research

Timely development of vaccines against sars-cov-2, Emerg. Microbes 395 Infec

Vaccines for sars-cov-2: lessons from other coronavirus 397 strains

In silico screening of 399 natural compounds against covid-19 by targeting mpro and ace2 using molecular 400 docking

In silico molecular 402 docking analysis for repurposing therapeutics against multiple proteins from 403 sars-cov-2

Curcumin, a traditional spice 406 component, can hold the promise against COVID-19?

Andrographolide and its 409 fluorescent derivative inhibit the main proteases of 2019-ncov and sars-cov 410 through covalent linkage

Research and development on 414 therapeutic agents and vaccines for COVID-19 and related human coronavirus 415 diseases

T. severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)

Insights into antiviral mechanisms 430 of remdesivir, lopinavir/ritonavir and chloroquine/hydroxychloroquine affecting 431 the new sars-cov-2

Remdesivir is a direct-acting antiviral that inhibits rna-dependent rna 434 polymerase from severe acute respiratory syndrome coronavirus 2 with high 435 potency

Anti-inflammatory activity of

H1-receptor antagonists: review of recent experimental research

Molecular docking and dynamics simulation of FDA approved drugs with the 441 main protease from 2019 novel coronavirus

Systemic in 445 silico screening in drug discovery for coronavirus disease (covid-19) with an 446 online interactive web server

Chloroquine and hydroxychloroquine as 452 ace2 blockers to inhibit viropexis of 2019-ncov spike pseudotyped virus

A vascular smooth muscle/cell 455 membrane chromatography-offline-gas chromatography/mass spectrometry 456 method for recognition, separation and identification of active components from 457 traditional chinese medicines

Coating and fusing cell membranes onto a silica 459 surface and their chromatographic characteristics

Identification of SARS-CoV-2 entry 472 inhibitors among already approved drugs

Pharmacology and clinical efficacy of desloratadine as an 475 anti-allergic and anti-inflammatory drug

Impact of desloratadine 477 and loratadine on the crosstalk between human keratinocytes and leukocytes: 478 implications for anti-inflammatory activity of antihistamines

Effect of desloratadine and loratadine on rhinovirus-induced intercellular 482 adhesion molecule 1 upregulation and promoter activation in respiratory 483 epithelial cells

Discovery of a novel coronavirus associated with the recent 486 pneumonia outbreak in humans and its potential bat origin

Structural basis of receptor recognition by 489 sars-cov-2

Structure of the SARS-CoV-2 spike receptor-binding domain 492 bound to the ACE2 receptor

Establishment 495 and validation of a pseudovirus neutralization assay for sars