PMC:7127386 / 6210-11786 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7127386","sourcedb":"PMC","sourceid":"7127386","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7127386","text":"Among the 16 compounds we tested, remdesivir, lopinavir, homoharringtonine, and emetine dihydrochloride were found to inhibit SARS-CoV-2 replication in Vero E6 cells with EC50 under 100 μM (Table 1 ). Importantly, we observed that some of the compounds currently undergoing clinical trials such as ribavirin, favipiravir, oseltamivir, or baloxavir showed no apparent antiviral effect against the SARS-CoV-2 virus in vitro at concentrations under 100 μM (Table 1). Remdesivir is a 1′-cyano-substituted adenosine analogue that has been shown to inhibit human coronaviruses (hCoV-OC43 and hCoV-229E) SARS-CoV, MERS-CoV, and SARS-CoV-2 (Brown et al., 2019; Sheahan et al., 2017; de Wit et al., 2020). It is currently evaluated in phase 4 clinical trials for SARS-CoV-2. A recent study fitted viral load in linear scale (eg. the percentage of inhibition) under increasing concentrations of remdesivir reported EC50 against SARS-CoV-2 virus at 0.77 μM (Wang et al., 2020). We fitted viral load in logarithm scale (log10TCID50/mL and log10 viral RNA copies/mL) under increasing concentration of remdesivir and determined EC50 at 23.15 μM and 26.90 μM, respectively (Fig. 1 A and Table 1). Two mutations (F476L and V553L) in the RNA-dependent RNA polymerase nsp12 of a murine hepatitis virus have been previously reported to confer resistance to remdesivir (Agostini et al., 2018). Due to insertions and deletions in nsp12, these two conserved residues are mapped at F480 and V557 in the SARS-CoV-2 isolate (GISAID# EPI_ISL_412028) used for the experiments, which should remain sensitive for remdesivir. Other adenosine analogues (galidesivir, tenofovor, or fludarabine phosphate) or nucleoside analogues (favipiravir, ribavirin, R-1479) did not inhibit viral replication under 100 μM (Table 1). However, nucleoside analogues require metabolic activation into their triphosphate forms by host cellular nucleoside kinases, which may differ among cell types. Further evaluation of the effect of nucleoside analogues in primary human airway epithelial cells would facilitate the interpretation of the results.\nTable 1 Antiviral activity of 16 compounds against SARS-CoV-2 in Vero E6 cells.\nCompounds Inhibition of SARS-CoV-2 in vitro, μM\nName Bioactivity Clinical application CAS No. CC50, μMa CPE inhibitionb Reduction in infectious virusc (EC50) Reduction in viral RNA copyd (EC50)\nRemdesivir adenosine analogue Phase 4 trials for treatment of Ebola or SARS-CoV-2 1809249-37-3 \u003e100 25 23.15 26.90\nFavipiravir guanineanalogue Approved in Japan and China for treatment of influenza infection 259793-96-9 \u003e100 \u003e100 \u003e100 \u003e100\nRibavirin guanosine analogue FDA approved for treatment of chronic hepatitis C infection 36791-04-5 \u003e100 500 \u003e500 \u003e500\nGalidesivir adenosine analogue Phase 2 trial for yellow fever virus infection 222631-44-9 \u003e100 100 \u003e100 \u003e100\nR-1479 cytidineanalogue Phase 2 trial for treatment of dengue virus infection 478182-28-4 \u003e100 \u003e100 N.D. N.D.\nTenofovor adenosine analogue FDA approved for treatment of HIV-1 and HBV 147127-20-6 \u003e100 \u003e100 N.D. N.D.\nFludarabine phosphate adenosine analogue FDA approved for treatment of B-cell chronic lymphocytic leukemia 75607-67-9 \u003e100 \u003e100 N.D. N.D.\nLopinavir protease inhibitor FDA approved for treatment of HIV-1 infection in combination with ritonavir 192725-17-0 49.75 25 26.63 26.10\nRitonavir protease inhibitor FDA approved for treatment of HIV-1 infection in combination with other antiretroviral agents 155213-67-5 48.91 \u003e100 \u003e100 \u003e100\nEmetine hydrochloride anti-protozoal Approved in China for severe invasive amoebiasis 316-42-7 56.46 1.5625 0.46 0.50\nOritavancin diphosphate antibiotics FDA approved treatment for skin infection caused by Gram positive bacteria 192564-14-0 N.D. \u003e100 N.D. N.D.\nDalbavancin hydrochloride antibiotics FDA approved treatment for skin infection caused by Gram positive bacteria 2227366-51-8 N.D. \u003e100 N.D. N.D.\nHomoharringtonine anti-cancer FDA approved treatment for chronic myeloid leukemia 26833-87-4 59.75 3.125 2.55 2.14\nOseltamivir carboxylate antiviral, neuraminidase inhibitor FDA approved treatment for influenza infection 187227-45-8 \u003e100 \u003e100 \u003e100 \u003e100\nBaloxivir acid antiviral, endonuclease inhibitor FDA approved treatment for influenza infection 1985605-59-1 85.90 \u003e100 \u003e100 \u003e100\nChlorpromazine hydrochloride antagonist for post-synaptic receptors FDA approved treatment for schizophrenia 69-09-0 21.29 \u003e100 N.D. N.D.\nN.D. Not determined.\na CC50 was determined with serially-diluted compounds in Vero E6 cells at 48 h post-incubation using CellTiter-Glow Luminescent Cell Viability Assay (Promega).\nb Compounds were serially 2-fold or 4-fold diluted from 100 μM, except ribavirin which was started at 500 μM. Cytopathic effects (CPE) of SARS-CoV-2 virus in Vero E6 cells under increasing concentration of the compounds were observed at 48 h post-infection. The lowest concentration of the compound with 100% CPE inhibition (eg. exhibiting comparable CPE of non-infected controls) was recorded.\nc EC50 determined by infectious virus yield in culture supernatant at 48h post-infection (log10 TCID50/mL).\nd EC50 determined by viral RNA copy numbers in culture supernatant at 48h post-infection (log10 RNA copies/mL).\nFig. 1 Antiviral activity of remdesivir (A), lopinavir (B), homorringtonine (C) and emetine dihydrochloride (D) against SARS-CoV-2 virus in vitro. Infectious viral loads (log10TCID50/mL left Y axis) and viability (normalized to the ATP level of the Vero E6 cells incubated with infection media) under increasing concentrations of the antiviral compounds are 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