Remdesivir and EIDD-1931 interact with human equilibrative nucleoside transporters 1 and 2: Implications for reaching SARS-CoV-2 viral sanctuary sites

Siennah R. Miller, Meghan E. McGrath, Kimberley M. Zorn, Sean Ekins, Stephen H. Wright, Nathan J. Cherrington

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Equilibrative nucleoside transporters (ENTs) are present at the blood-testis barrier (BTB), where they can facilitate antiviral drug disposition to eliminate a sanctuary site for viruses detectable in semen. The purpose of this study was to investigate ENT-drug interactions with three nucleoside analogs, remdesivir, molnupiravir, and molnupiravir’s active metabolite, b-D-N4hydroxycytidine (EIDD-1931), and four non-nucleoside molecules repurposed as antivirals for coronavirus disease 2019 (COVID-19). The study used three-dimensional pharmacophores for ENT1 and ENT2 substrates and inhibitors and Bayesian machine learning models to identify potential interactions with these transporters. In vitro transport experiments demonstrated that remdesivir was the most potent inhibitor of ENT-mediated [3H]uridine uptake (ENT1 IC50: 39 mM; ENT2 IC50: 77 mM), followed by EIDD-1931 (ENT1 IC50: 259 mM; ENT2 IC50: 467 mM), whereas molnupiravir was a modest inhibitor (ENT1 IC50: 701 mM; ENT2 IC50: 851 mM). Other proposed antivirals failed to inhibit ENT-mediated [3H]uridine uptake below 1 mM. Remdesivir accumulation decreased in the presence of 6-S-[(4-nitrophenyl)methyl]-6-thioinosine (NBMPR) by 30% in ENT1 cells (P 5 0.0248) and 27% in ENT2 cells (P 5 0.0054). EIDD-1931 accumulation decreased in the presence of NBMPR by 77% in ENT1 cells (P 5 0.0463) and by 64% in ENT2 cells (P 5 0.0132), which supported computational predictions that both are ENT substrates that may be important for efficacy against COVID-19. NBMPR failed to decrease molnupiravir uptake, suggesting that ENT interaction is likely inhibitory. Our combined computational and in vitro data can be used to identify additional ENT-drug interactions to improve our understanding of drugs that can circumvent the BTB.

Original languageEnglish (US)
Pages (from-to)548-557
Number of pages10
JournalMolecular pharmacology
Volume100
Issue number6
DOIs
StatePublished - Dec 1 2021

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

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