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Related Experiment Video

Updated: Oct 31, 2025

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Probing the SAM Binding Site of SARS-CoV-2 Nsp14 In Vitro Using SAM Competitive Inhibitors Guides Developing

Kanchan Devkota1, Matthieu Schapira1,2, Sumera Perveen1

  • 1Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada.

SLAS Discovery : Advancing Life Sciences R & D
|July 1, 2021
PubMed
Summary
This summary is machine-generated.

Researchers screened compounds to inhibit SARS-CoV-2 nonstructural protein 14 (nsp14) methyltransferase activity, crucial for viral RNA capping. Two compounds, SS148 and DS0464, showed potent inhibition and selectivity, offering potential for new COVID-19 antivirals.

Keywords:
COVID-19SARS-CoV-2coronavirusnsp14

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Area of Science:

  • Virology
  • Biochemistry
  • Drug Discovery

Background:

  • The COVID-19 pandemic highlights the urgent need for effective antiviral therapies.
  • SARS-CoV-2 utilizes RNA capping, involving nonstructural protein 14 (nsp14) methyltransferase (MTase) activity, to evade the host immune system.
  • Targeting nsp14 MTase offers a potential strategy for developing novel antivirals against coronaviruses.

Purpose of the Study:

  • To identify small-molecule inhibitors targeting the methyltransferase activity of SARS-CoV-2 nsp14.
  • To evaluate the inhibitory potency and selectivity of identified compounds against nsp14.
  • To explore structure-activity relationships for optimizing nsp14 inhibitors.

Main Methods:

  • Development and application of a radiometric methyltransferase (MTase) assay.
  • Screening of a library of 161 synthesized S-adenosylmethionine (SAM) competitive inhibitors and SAM analogs.
  • Evaluation of compound selectivity against human protein lysine methyltransferases and other MTases.

Main Results:

  • Six screening hits were identified, with SS148 inhibiting nsp14 MTase activity (IC50 = 70 ± 6 nM) and showing selectivity against human protein lysine MTases.
  • DS0464 demonstrated a bisubstrate competitive inhibition mechanism (IC50 = 1.1 ± 0.2 µM) and selectivity against a broad range of RNA, DNA, and protein MTases.
  • Structure-activity relationship insights were gained for optimizing nsp14 inhibitors.

Conclusions:

  • The identified compounds, particularly SS148 and DS0464, represent promising leads for developing selective nsp14 inhibitors.
  • These findings pave the way for a novel class of antiviral agents targeting SARS-CoV-2 and potentially other coronaviruses.
  • Further optimization of these compounds could lead to effective therapeutics against COVID-19.