Resistance Studies of a Dithiazol Analogue, DBPR110, as a Potential Hepatitis C Virus NS5A Inhibitor in Replicon Systems

• DOI: 10.1128/aac.01403-12
• Publication Date: 2012-11-20T05:25:19Z
• Authors: Hui-Mei Lin; Jing-Chyi Wang; Han-Shu Hu; Pei-Shan Wu; Wen-Hung Wang; Su-Ying Wu; Chi-Chen Yang; Teng-Kuang Yeh; Tsu-An Hsu; Weir-Torn Jiaang; Yu-Sheng Chao; Jyh-Haur Chern; Andrew Yueh
• Keywords: 0301 basic medicine; Pyrrolidines; Genotype; Sequence Analysis, RNA; Interferon-alpha; Hepacivirus; Hepatitis C, Chronic; Viral Nonstructural Proteins; Antiviral Agents; 3. Good health; Thiazoles; 03 medical and health sciences; Amino Acid Substitution; Cell Line, Tumor; Drug Resistance, Viral; Mutation; Humans; RNA, Viral; Replicon; Protein Binding

ABSTRACTHepatitis C virus (HCV), a member of theFlaviviridaefamily, affects approximately 3% of the world's population and is becoming the leading cause of liver disease in the world. Therefore, the development of novel or more effective treatment strategies to treat chronic HCV infection is urgently needed. In our previous study, we identified a potential HCV NS5A inhibitor, BP008. After further systemic optimization, we discovered a more potent HCV inhibitor, DBPR110. DBPR110 reduced the reporter expression of the HCV1b replicon with a 50% effective concentration (EC50) and a selective index value of 3.9 ± 0.9 pM and >12,800,000, respectively. DBPR110 reduced HCV2a replicon activity with an EC50and a selective index value of 228.8 ± 98.4 pM and >173,130, respectively. Sequencing analyses of several individual clones derived from the DBPR110-resistant RNAs purified from cells harboring genotype 1b and 2a HCV replicons revealed that amino acid substitutions mainly within the N-terminal region (domain I) of NS5A were associated with decreased inhibitor susceptibility. P58L/T and Y93H/N in genotype 1b and T24A, P58L, and Y93H in the genotype 2a replicon were the key substitutions for resistance selection. In the 1b replicon, V153M, M202L, and M265V play a compensatory role in replication and drug resistance. Moreover, DBPR110 displayed synergistic effects with alpha interferon (IFN-α), an NS3 protease inhibitor, and an NS5B polymerase inhibitor. In summary, our results present an effective small-molecule inhibitor, DBPR110, that potentially targets HCV NS5A. DBPR110 could be part of a more effective therapeutic strategy for HCV in the future.