In Silico and In Vitro Evaluation of Some Amidine Derivatives as Hit Compounds towards Development of Inhibitors against Coronavirus Diseases

• Published in: Viruses Vol. 15; no. 5; p. 1171
• Main Authors: Hassan, Ahmed H E, El-Sayed, Selwan M, Yamamoto, Mizuki, Gohda, Jin, Matsumoto, Takehisa, Shirouzu, Mikako, Inoue, Jun-Ichiro, Kawaguchi, Yasushi, Mansour, Reem M A, Anvari, Abtin, Farahat, Abdelbasset A
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 15.05.2023; MDPI
• Subjects: Amidines; Antiviral agents; Antiviral Agents - chemistry; Antiviral Agents - pharmacology; Benzamidines - pharmacology; Cells; Coronavirus infections; Coronaviruses; COVID-19; COVID-19 vaccines; Drug therapy; Enzymes; Epidemics; Genomes; Health aspects; Humans; Infections; Influenza; Influenza A; Membrane fusion; Middle East respiratory syndrome; Middle East Respiratory Syndrome Coronavirus; Mutation; Pandemics; Pentamidine; Pharmacology, Experimental; Plasmids; Product development; Proteinase inhibitors; Proteins; Respiratory diseases; RNA polymerase; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; Testing; Thrombin; viral entry; Viral infections; Virus Internalization; Viruses; Egypt; United States > US; China; Germany; viral entry; antiviral agents; coronaviruses
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v15051171

Coronaviruses, including SARS-CoV-2, SARS-CoV, MERS-CoV and influenza A virus, require the host proteases to mediate viral entry into cells. Rather than targeting the continuously mutating viral proteins, targeting the conserved host-based entry mechanism could offer advantages. Nafamostat and camostat were discovered as covalent inhibitors of TMPRSS2 protease involved in viral entry. To circumvent their limitations, a reversible inhibitor might be required. Considering nafamostat structure and using pentamidine as a starting point, a small set of structurally diverse rigid analogues were designed and evaluated in silico to guide selection of compounds to be prepared for biological evaluation. Based on the results of in silico study, six compounds were prepared and evaluated in vitro. At the enzyme level, compounds - triggered potential TMPRSS2 inhibition with low micromolar IC concentrations, but they were less effective in cellular assays. Meanwhile, compound did not trigger potential TMPRSS2 inhibition at the enzyme level, but it showed potential cellular activity regarding inhibition of membrane fusion with a low micromolar IC value of 10.87 µM, suggesting its action could be mediated by another molecular target. Furthermore, in vitro evaluation showed that compound inhibited pseudovirus entry as well as thrombin and factor Xa. Together, this study presents compound as a hit compound that might serve as a starting point for developing potential viral entry inhibitors with possible application against coronaviruses.