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Discovering novel anti-HCV compounds with inhibitory activities toward HCV NS3/4A protease.

Yu Y, Jing JF, Tong XK, He PL, Li YC, Hu YH, Tang W, Zuo JP - Acta Pharmacol. Sin. (2014)

Bottom Line: Among them, two representative compounds HZ-1157 and LZ-110618-6 inhibited HCV NS3/4A protease with IC50 values of 1.0 and 0.68 μmol/L, respectively.Furthermore, HZ-1157 and LZ-110618-6 inhibited HCV infection in vitro with IC50 values of 0.82 and 0.11 μmol/L, respectively.Some 2,4-diaminoquinazoline derivatives and carboxamide analogues have been identified as novel anti-HCV compounds.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.

ABSTRACT

Aim: To discover novel hepatitis C virus (HCV) inhibitors and elucidate the mechanism of action of the active compounds.

Methods: HCV subgenomic replicon-based luciferase reporter cell line was used to screen 1200 synthetic compounds with novel structures. Huh7.5.1 cell line stably transfected with HCV NS3/4A protease reporter was established to investigate the anti-HCV mechanism of the active compounds. The active compounds were further examined in an in vitro HCV infection assay to confirm their anti-HCV activity.

Results: After two-round screening in the anti-HCV replicon assay, some 2,4-diaminoquinazoline derivatives and carboxamide analogues were found to possess anti-HCV replicon activities (the IC50 values were less than 5 μmol/L). Among them, two representative compounds HZ-1157 and LZ-110618-6 inhibited HCV NS3/4A protease with IC50 values of 1.0 and 0.68 μmol/L, respectively. Furthermore, HZ-1157 and LZ-110618-6 inhibited HCV infection in vitro with IC50 values of 0.82 and 0.11 μmol/L, respectively.

Conclusion: Some 2,4-diaminoquinazoline derivatives and carboxamide analogues have been identified as novel anti-HCV compounds.

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Related in: MedlinePlus

The reporter activities of HCV NS3/4A-Seap stably transfected clones. Huh7.5.1 cells were transfected with the pSelect-NS3/4-Seap plasmid using Lipofectamine 2000 transfection reagent and selected with 500 mg/L zeocin for 4 weeks. Cell colonies were picked and propagated to the proper confluency, and the Seap activities in culture supernatants were assayed.
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fig5: The reporter activities of HCV NS3/4A-Seap stably transfected clones. Huh7.5.1 cells were transfected with the pSelect-NS3/4-Seap plasmid using Lipofectamine 2000 transfection reagent and selected with 500 mg/L zeocin for 4 weeks. Cell colonies were picked and propagated to the proper confluency, and the Seap activities in culture supernatants were assayed.

Mentions: To establish a stably transfected HCV NS3/4A-Seap reporter cell line, Huh7.5.1 cells were transfected with pSelect-NS3/4A-Seap and selected by 500 mg/L zeocin for four weeks. We obtained five colonies of cells. The Seap levels in supernatants were tested, and clone 3, which had the highest Seap activity, was chosen and propagated (Figure 5). The inhibitory efficacy of telaprevir was tested in clone 3. The results showed that the IC50 of telaprevir was 0.931 μmol/L (Figure 6A). The CC50 was higher than 10 μmol/L (Figure 6B).


Discovering novel anti-HCV compounds with inhibitory activities toward HCV NS3/4A protease.

Yu Y, Jing JF, Tong XK, He PL, Li YC, Hu YH, Tang W, Zuo JP - Acta Pharmacol. Sin. (2014)

The reporter activities of HCV NS3/4A-Seap stably transfected clones. Huh7.5.1 cells were transfected with the pSelect-NS3/4-Seap plasmid using Lipofectamine 2000 transfection reagent and selected with 500 mg/L zeocin for 4 weeks. Cell colonies were picked and propagated to the proper confluency, and the Seap activities in culture supernatants were assayed.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4125721&req=5

fig5: The reporter activities of HCV NS3/4A-Seap stably transfected clones. Huh7.5.1 cells were transfected with the pSelect-NS3/4-Seap plasmid using Lipofectamine 2000 transfection reagent and selected with 500 mg/L zeocin for 4 weeks. Cell colonies were picked and propagated to the proper confluency, and the Seap activities in culture supernatants were assayed.
Mentions: To establish a stably transfected HCV NS3/4A-Seap reporter cell line, Huh7.5.1 cells were transfected with pSelect-NS3/4A-Seap and selected by 500 mg/L zeocin for four weeks. We obtained five colonies of cells. The Seap levels in supernatants were tested, and clone 3, which had the highest Seap activity, was chosen and propagated (Figure 5). The inhibitory efficacy of telaprevir was tested in clone 3. The results showed that the IC50 of telaprevir was 0.931 μmol/L (Figure 6A). The CC50 was higher than 10 μmol/L (Figure 6B).

Bottom Line: Among them, two representative compounds HZ-1157 and LZ-110618-6 inhibited HCV NS3/4A protease with IC50 values of 1.0 and 0.68 μmol/L, respectively.Furthermore, HZ-1157 and LZ-110618-6 inhibited HCV infection in vitro with IC50 values of 0.82 and 0.11 μmol/L, respectively.Some 2,4-diaminoquinazoline derivatives and carboxamide analogues have been identified as novel anti-HCV compounds.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.

ABSTRACT

Aim: To discover novel hepatitis C virus (HCV) inhibitors and elucidate the mechanism of action of the active compounds.

Methods: HCV subgenomic replicon-based luciferase reporter cell line was used to screen 1200 synthetic compounds with novel structures. Huh7.5.1 cell line stably transfected with HCV NS3/4A protease reporter was established to investigate the anti-HCV mechanism of the active compounds. The active compounds were further examined in an in vitro HCV infection assay to confirm their anti-HCV activity.

Results: After two-round screening in the anti-HCV replicon assay, some 2,4-diaminoquinazoline derivatives and carboxamide analogues were found to possess anti-HCV replicon activities (the IC50 values were less than 5 μmol/L). Among them, two representative compounds HZ-1157 and LZ-110618-6 inhibited HCV NS3/4A protease with IC50 values of 1.0 and 0.68 μmol/L, respectively. Furthermore, HZ-1157 and LZ-110618-6 inhibited HCV infection in vitro with IC50 values of 0.82 and 0.11 μmol/L, respectively.

Conclusion: Some 2,4-diaminoquinazoline derivatives and carboxamide analogues have been identified as novel anti-HCV compounds.

Show MeSH
Related in: MedlinePlus