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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 screening results of the anti-HCV replicon assay. 1200 synthetic compounds with novel structure were screened by using the HCV subgenomic replicon-based reporter cell line (J399LM, genotype 2a). Compounds were co-cultured with cells for 48 h. The luciferase signal was read and compared to control wells without compounds. Compounds with reporter signal lower than the mean-3xSD (lower dashed line) of control wells (without compound) were considered to be active compounds and selected for further tests. Normal Huh7.5.1 cells served as the blank.
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fig2: The screening results of the anti-HCV replicon assay. 1200 synthetic compounds with novel structure were screened by using the HCV subgenomic replicon-based reporter cell line (J399LM, genotype 2a). Compounds were co-cultured with cells for 48 h. The luciferase signal was read and compared to control wells without compounds. Compounds with reporter signal lower than the mean-3xSD (lower dashed line) of control wells (without compound) were considered to be active compounds and selected for further tests. Normal Huh7.5.1 cells served as the blank.

Mentions: In total, 1200 synthetic compounds were screened in a HCV replicon reporter system, which was a Huh7 harboring a HCV subgenome with a Renilla luciferase reporter. The results of the preliminary screening are shown in Figure 2, with the luciferase signal of each compound plotted. Of the 1200 compounds screened, those with luciferase signals lower than the threshold (Mean-3×SD of the control well, lower dashed line, Figure 2) were considered to be anti-HCV active compounds and selected for the second-round test. In the second-round test, candidate compounds were serially diluted and tested by the same HCV replicon assay. IC50 values of the selected compounds are shown in Table 1. We discovered two categories of compounds, 2,4-diaminoquinazoline derivatives and carboxamide analogues, that exhibited anti-HCV activity at reasonable concentrations (IC50<5 μmol/L). These compounds were selected for further investigation of their anti-HCV mechanisms.


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 screening results of the anti-HCV replicon assay. 1200 synthetic compounds with novel structure were screened by using the HCV subgenomic replicon-based reporter cell line (J399LM, genotype 2a). Compounds were co-cultured with cells for 48 h. The luciferase signal was read and compared to control wells without compounds. Compounds with reporter signal lower than the mean-3xSD (lower dashed line) of control wells (without compound) were considered to be active compounds and selected for further tests. Normal Huh7.5.1 cells served as the blank.
© Copyright Policy
Related In: Results  -  Collection

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

fig2: The screening results of the anti-HCV replicon assay. 1200 synthetic compounds with novel structure were screened by using the HCV subgenomic replicon-based reporter cell line (J399LM, genotype 2a). Compounds were co-cultured with cells for 48 h. The luciferase signal was read and compared to control wells without compounds. Compounds with reporter signal lower than the mean-3xSD (lower dashed line) of control wells (without compound) were considered to be active compounds and selected for further tests. Normal Huh7.5.1 cells served as the blank.
Mentions: In total, 1200 synthetic compounds were screened in a HCV replicon reporter system, which was a Huh7 harboring a HCV subgenome with a Renilla luciferase reporter. The results of the preliminary screening are shown in Figure 2, with the luciferase signal of each compound plotted. Of the 1200 compounds screened, those with luciferase signals lower than the threshold (Mean-3×SD of the control well, lower dashed line, Figure 2) were considered to be anti-HCV active compounds and selected for the second-round test. In the second-round test, candidate compounds were serially diluted and tested by the same HCV replicon assay. IC50 values of the selected compounds are shown in Table 1. We discovered two categories of compounds, 2,4-diaminoquinazoline derivatives and carboxamide analogues, that exhibited anti-HCV activity at reasonable concentrations (IC50<5 μmol/L). These compounds were selected for further investigation of their anti-HCV mechanisms.

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