PMC:7291971 / 22531-25169 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7291971","sourcedb":"PMC","sourceid":"7291971","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7291971","text":"2.4 Dose-response testing of selected compounds against SARS-CoV nsp14 and hRNMT\nTo confirm the observed inhibition of some adenine dinucleosides against SARS-CoV nsp14 N7-MTase, we further tested 10 compounds in a dose-response assay (Table 2 , Supporting Information Fig. S2). The selection of these compounds was based on their percentage inhibition higher than 50% against SARS-CoV nsp14 (Table 1). After pre-incubation with increasing concentrations of dinucleosides, the MTase activity was measured by FBA. The IC50 of compounds, deduced from Hill slope equation (Y = 100/(1+((X/IC50)^Hillslope) curve-fitting, ranged from 0.6 μM to 70.4 μM on SARS-CoV nsp14 and 10.9 μM to \u003e 500 μM on hRNMT activity. Among all the potential inhibitors, three compounds 2, 6 and 11 showed the lowest inhibitory activity on SARS-CoV nsp14 with IC50 \u003e 40 μM, the others have IC50 values in the micromolar range except 13 that showed an IC50 value in the submicromolar range. The potency of the derivative 13 to inhibit SARS-CoV nsp14 was comparable to the broad spectrum inhibitor sinefungin. However, it is noteworthy that compound 13 specifically inhibits the N7-MTase of SARS-CoV with a high 413-fold of specificity in dose-response testing whereas sinefungin is also active against hRNMT [11]. Remarkably 13 bearing a chlorine atom in “para” position and a nitro group in “meta” position displayed the best inhibition which was 6.5–9.5 fold more potent than the derivatives 10 and 12, respectively that contain the nitro in “ortho” position and a lipophilic group CF3 or MeO in “para” position. The comparison of IC50 values of 14 and 13 indicates that the addition of a nitro group in the phenyl ring increased the inhibitory activity by 2.5 fold in 13. These results confirm that the chlorine atom in “para” position and the presence of a nitro group seem crucial for submicromolar SARS-CoV nsp14 inhibitory activity.\nTable 2 Comparison of IC50 values of 10 most active compounds on SARS-CoV nsp14 and human RNMT activities.\nCompounds SARS-CoV nsp14 IC50a (μM) hRNMTIC50 (μM)\nSinefunginb 0.36 \u003c0.05\n2 40.6 ± 3.2 10.9 ± 1.0\n6 55.5 ± 5.1 \u003e500\n9 2.6 ± 0.2 \u003e500\n10 3.9 ± 0.4 \u003e500\n11 70.4 ± 4.9 169.3 ± 30.4\n12 5.7 ± 0.5 275.9 ± 28.7\n13 0.6 ± 0.1 247.5 ± 14.9\n14 1.5 ± 0.1 n.d\n15 2.4 ± 0.2 n.i\n16 9.9 ± 0.9 n.d\na Concentration inhibiting MTase activity by 50%; mean value from three independent experiments.\nb values from the literature [11]. The MTase activity for IC50 determinations was measured using a filter binding assay as described above. n.i: no inhibition detected at 50 μM; n.d: not determined due to inhibition lower than 50% at 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