The schematic diagram of HCV NS3/4A-Seap expression pla

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. Show MeSH Major Antiviral Agents/chemistry* Hepacivirus/drug effects/metabolism Hepatitis C/drug therapy/virology Quinazolines/chemistry/pharmacology Viral Nonstructural Proteins/antagonists & inhibitors/metabolism Minor* Amides/chemistry/pharmacology Cell Line Drug Evaluation, Preclinical Humans Related in: MedlinePlus	© Copyright Policy Related In: Results - Collection Show All Figures getmorefigures.php?uid=PMC4125721&req=5 fig1: The schematic diagram of HCV NS3/4A-Seap expression plasmids. The Seap gene with an N-terminal signal was fused in-frame to the HCV NS3/4A coding sequence. NS3/4A and Seap amino acids in the fusion regions are shown by capitalized characters. HDS is the catalytic triad of the HCV NS3 protease. The S to A mutation in the catalytic triad results in a disabled NS3 protease. The NS4A/4B cleavage site is indicated by an arrow. Mentions: For the pSelect-NS3/4A-Seap plasmid, the NS3/4A coding sequence, including the cleavage site, was amplified from the full length JFH1 strain plasmid pJFH1T (genotype 2a) by PCR with the forward primer 5′-ccatggTTGCTCCCATCACTGCTT-3′ and the reverse primer 5′-ccatggGCATTCCTCCATCTCATCAA-3′ and cloned into the NcoI site of the pSELECT-zeo-Seap vector (InvivoGen, San Diego, California, USA). The schematic diagram of this plasmid is shown in Figure 1. For the pSelect-NS3/4A(mut)–Seap plasmid, the serine of the HDS catalytic triad was changed to alanine (S139A).

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.

Show MeSH

Related in: MedlinePlus

The schematic diagram of HCV NS3/4A-Seap expression plasmids. The Seap gene with an N-terminal signal was fused in-frame to the HCV NS3/4A coding sequence. NS3/4A and Seap amino acids in the fusion regions are shown by capitalized characters. HDS is the catalytic triad of the HCV NS3 protease. The S to A mutation in the catalytic triad results in a disabled NS3 protease. The NS4A/4B cleavage site is indicated by an arrow.

Related In: Results - Collection

Show All Figures

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fig1: The schematic diagram of HCV NS3/4A-Seap expression plasmids. The Seap gene with an N-terminal signal was fused in-frame to the HCV NS3/4A coding sequence. NS3/4A and Seap amino acids in the fusion regions are shown by capitalized characters. HDS is the catalytic triad of the HCV NS3 protease. The S to A mutation in the catalytic triad results in a disabled NS3 protease. The NS4A/4B cleavage site is indicated by an arrow.

Mentions: For the pSelect-NS3/4A-Seap plasmid, the NS3/4A coding sequence, including the cleavage site, was amplified from the full length JFH1 strain plasmid pJFH1T (genotype 2a) by PCR with the forward primer 5′-ccatggTTGCTCCCATCACTGCTT-3′ and the reverse primer 5′-ccatggGCATTCCTCCATCTCATCAA-3′ and cloned into the NcoI site of the pSELECT-zeo-Seap vector (InvivoGen, San Diego, California, USA). The schematic diagram of this plasmid is shown in Figure 1. For the pSelect-NS3/4A(mut)–Seap plasmid, the serine of the HDS catalytic triad was changed to alanine (S139A).