Probing the SAM Binding Site of SARS-CoV-2 Nsp14 In Vitro Using SAM Competitive Inhibitors Guides Developing Selective Bisubstrate Inhibitors
| Autor: | Peter Loppnau, Minkui Luo, Pegah Ghiabi, Albina Bolotokova, Irene Chau, Fengling Li, Jing Liu, Matthieu Schapira, Karla J. F. Satchell, Sumera Perveen, Ke Wang, Aliakbar Khalili Yazdi, Paul V. Fish, Jian Jin, Peter Brown, Deyao Li, Masoud Vedadi, David Smil, Kanchan Devkota, Taraneh Hajian |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
RNA capping
Methyltransferase coronavirus Viral Nonstructural Proteins medicine.disease_cause Virus Replication 01 natural sciences Biochemistry Antiviral Agents Methylation Virus Article Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound medicine Humans Binding site Pandemics 030304 developmental biology Coronavirus Original Research 0303 health sciences Binding Sites SARS-CoV-2 RNA COVID-19 Small molecule 0104 chemical sciences 010404 medicinal & biomolecular chemistry chemistry Exoribonucleases nsp14 Molecular Medicine RNA Viral DNA Biotechnology Protein Binding |
| Zdroj: | Slas Discovery bioRxiv |
| ISSN: | 2472-5560 2472-5552 |
| Popis: | The COVID-19 pandemic has clearly brought the healthcare systems world-wide to a breaking point along with devastating socioeconomic consequences. The SARS-CoV-2 virus which causes the disease uses RNA capping to evade the human immune system. Non-structural protein (nsp) 14 is one of the 16 nsps in SARS-CoV-2 and catalyzes the methylation of the viral RNA at N7-guanosine in the cap formation process. To discover small molecule inhibitors of nsp14 methyltransferase (MT) activity, we developed and employed a radiometric MT assay to screen a library of 161 in house synthesized S-adenosylmethionine (SAM) competitive methyltransferase inhibitors and SAM analogs. Among seven identified screening hits, SS148 inhibited nsp14 MT activity with an IC50value of 70 ± 6 nM and was selective against 20 human protein lysine methyltransferases indicating significant differences in SAM binding sites. Interestingly, DS0464 with IC50value of 1.1 ± 0.2 μM showed a bi-substrate competitive inhibitor mechanism of action. Modeling the binding of this compound to nsp14 suggests that the terminal phenyl group extends into the RNA binding site. DS0464 was also selective against 28 out of 33 RNA, DNA, and protein methyltransferases. The structure-activity relationship provided by these compounds should guide the optimization of selective bi-substrate nsp14 inhibitors and may provide a path towards a novel class of antivirals against COVID-19, and possibly other coronaviruses. |
| Databáze: | OpenAIRE |
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