CETSA MS Profiling for a Comparative Assessment of FDA-Approved Antivirals Repurposed for COVID-19 Therapy Identifies TRIP13 as a Remdesivir Off-Target

Abstract

The reuse of preexisting small molecules for a novel emerging disease threat is a rapid measure to discover unknown applications for previously validated therapies. A pertinent and recent example where such a strategy could be employed is in the fight against coronavirus disease 2019 (COVID-19). Therapies designed or discovered to target viral proteins also have off-target effects on the host proteome when employed in a complex physiological environment. This study aims to assess these host cell targets for a panel of FDA-approved antiviral compounds including remdesivir, using the cellular thermal shift assay (CETSA) coupled with mass spectrometry (CETSA MS) in noninfected cells. CETSA MS is a powerful method to delineate direct and indirect interactions between small molecules and protein targets in intact cells. Biologically active compounds can induce changes in thermal stability, in their primary binding partners, and in proteins that in turn interact with the direct targets. Such engagement of host targets by antiviral drugs may contribute to the clinical effect against the virus but can also constitute a liability. We present here a comparative study of CETSA molecular target engagement fingerprints of antiviral drugs to better understand the link between off-targets and efficacy.

Comprehensive Summary Supporting Clinical Investigation of GS-441524 for Covid-19 Treatment

“…Some have suggested that the initial phosphorylation reaction of GS-441524 occurs inefficiently, pointing to the rate-limiting phosphorylation of the sofosbuvir nucleoside (PSI-6206) by the enzyme deoxycytidine kinase (dCK) 9 . However, a recent study by Friman and colleagues 44 have demonstrated that, at least in the HepG2 cell line, GS-441524 is not a substrate of dCK (Figure 14). These data further suggest that ADK is likely the main phosphorylating enzyme for GS-441524.…”

A Comprehensive Review of Drug Repurposing Strategies against Known Drug Targets of COVID-19

“…The experiment was designed to study off-target effects of remdesivir and chloroquine as repurposed drugs for targeting SARS-CoV-2. This leads to a hopeful venture to further improve or develop fortuitous therapies for SARS-CoV-2 infection [34].…”

Development of Extracellular Vesicle Therapeutics: Challenges, Considerations, and Opportunities

“…The application of multi-omic technologies, such as transcriptomics and proteomics are emerging as crucial for understanding how EVs perform therapeutic functions ( Li et al, 2021d ). In the wider drug development field, omics technologies are already in use to better understand mechanisms of action and identify off-target effects ( Jia et al, 2019 ; Friman et al, 2021 ). Additionally, comprehensive profiling of patient and population proteomes/transcriptomes contribute to in silico predictions of drug efficacy [reviewed in El-Khateeb et al (2019) ].…”