Article Figures & Data
Figures
- FIG. 1.
Chemical structures of various nucleosides and antimetabolites.
- FIG. 2.
Dynamics of cell growth and HCV RNA levels after exposure to control anti-HCV compounds. HCV replicon cells were seeded at approximately 104 cells per well in a 24-well plate. Over a 7-day period, cells were counted daily, and rRNA and HCV RNA were quantified by quantitative reverse transcription-PCR. (A) IFN-α-2a at 100 IU/ml. (B) Ribavirin at 100 μM. (C) 2′-C-CH3-C at 100 μM. (D) 2′-C-CH3-A at 20 μM. •, cell proliferation in the absence of the compound; ○, cell proliferation in the presence of the compound; ▾, HCV RNA levels in untreated cells; ▿, HCV RNA levels in the presence of the compound. The points shown are averages ± standard deviations (S.D.) (error bars) for three independent experiments.
- FIG. 3.
Dynamics of the cell growth and HCV RNA levels after exposure to selected antimetabolites. The experimental method used is described in the legend to Fig. 2. (A) PALA at 10 μM. (B) PZF at 5 μM. (C) CP-C at 25 μM. (D) CPE-C at 2.5 μM. •, cell proliferation in the absence of the compound; ○, cell proliferation in the presence of the compound; ▾, HCV RNA levels in untreated cells; ▿, HCV RNA levels in the presence of the compound. The points are averages ± standard deviations (S.D.) (error bars) for three independent experiments.
- FIG. 4.
Replicon RNA changes per cell. The plots were obtained from collected data as shown in Fig. 2 and 3, in which log10 changes for cell count and replicon RNA levels were subtracted from each other. (A) Control antiviral compounds. •, no drug control; ▾, IFN-α-2a at 100 IU/ml; ▪, ribavirin at 100 μM; ⧫, 2′-C-CH3-C at 100 μM; ▴, 2′-C-CH3-A at 20 μM. (B) Selection of the most important antimetabolites. •, no drug control; ▾, dFdC at 50 nM; ▪, CP-C at 25 μM; ⧫, CPE-C at 2.5 μM; ▴, PALA at 10 μM; ⬢, PZF at 5 μM.
Tables
- TABLE 1.
Antiviral and cytotoxicity effects of antimetabolites on HCV replicon-transfected Huh-7 cells
Compound log10 RNA reductiona at 100 μM for: Corrected HCV RNA HCV rRNA log10 reductiona at 100 μM log10 reduction at 10 μM EC90, μM IMPDH inhibitors (EC 1.1.1.205) Mizoribine 0.29 ± 0.74 0.21 ± 0.50 0.08 ± 0.82 −0.14 ± 0.12 >100 Tiazofurin 0.86 ± 0.27 0.99 ± 0.35 −0.13 ± 0.37 0.04 ± 0.10 >100 Mycophenolic acid 1.15 ± 0.43 1.09 ± 0.28 0.07 ± 0.47 0.22 ± 0.01 >100 C2-MAD 1.09 ± 0.21 1.00 ± 0.15 0.08 ± 0.24 0.36 ± 0.21 >100 Ribonucleotide reductase inhibitors (EC 1.17.4.1; EC 1.17.4.2) Guanazole 0.25 ± 0.11 0.07 ± 0.03 0.32 ± 0.08 0.05 ± 0.08 >100 Hydroxyurea 0.17 ± 0.08 0.25 ± 0.20 −0.08 ± 0.16 0.06 ± 0.04 >100 Tezacytabine 1.59 ± 0.08 1.78 ± 0.69 −0.19 ± 0.49 0.63 ± 0.07 >100 Deferoxamine 1.00 ± 0.06 0.92 ± 0.08 0.08 ± 0.03 0.17 ± 0.11 >100 CTP synthase inhibitors (E.C. 6.3.4.2.) CP-C 1.97 ± 0.38 0.91 ± 0.13 1.06 ± 0.26 0.64 ± 0.10 25 CPE-C 2.47 ± 0.33 1.21 ± 0.16 1.26 ± 0.51 1.43 ± 0.01 2.5 3-DU 1.41 ± 0.09 0.48 ± 0.11 0.94 ± 0.20 0.13 ± 0.10 ∼100 dFdC 1.87 ± 0.16 0.59 ± 0.05 1.29 ± 0.11 1.32 ± 0.08 Too toxic OMPDC inhibitors (EC 4.1.1.23) 6-Azauridine 0.25 ± 0.09 0.61 ± 0.18 −0.36 ± 0.16 0.12 ± 0.05 >100 2-Thio-6-azauridine 0.16 ± 0.04 −0.02 ± 0.12 0.19 ± 0.09 0.12 ± 0.10 >100 PZF 1.88 ± 0.05 0.42 ± 0.03 1.46 ± 0.08 1.16 ± 0.21 3.80 ATC inhibitors (EC 2.1.3.2) PALA 1.77 ± 0.02 0.48 ± 0.02 1.30 ± 0.05 1.18 ± 0.11 7.60 Dihydroorotate dehydrogenase inhibitors (E.C. 1.3.3.1) Brequinar (NSC-368390) −0.05 ± 0.05 0.29 ± 0.01 −0.34 ± 0.04 −0.17 ± 0.09 >100 Dichloroallyl lawsone (NSC-126771) 1.27 ± 0.02 2.13 ± 0.15 −0.86 ± 0.17 −0.52 ± 0.01 >100 Thymidylate synthase inhibitors (E.C. 2.1.1.45) 2′-Deoxy-5-fluorouridine 0.76 ± 0.06 0.73 ± 0.35 0.04 ± 0.25 0.23 ± 0.05 >100 Methotrexate 0.18 ± 0.01 0.07 ± 0.10 0.11 ± 0.09 0.15 ± 0.01 >100 Controls IFN-α-2a 1.57 ± 0.26 0.21 ± 0.21 1.36 ± 0.37 NAb 4.5 IU/ml Ribavirin 1.96 ± 0.28 0.91 ± 0.12 1.05 ± 0.29 0.16 ± 0.10 ∼100 2′-C-CH3-A 2.32 ± 0.11 2.96 ± 0.08 −0.64 ± 0.18 2.05 1.4 2′-C-CH3-C 2.20 ± 0.52 −0.02 ± 0.05 2.21 ± 0.47 1.0 10.4
