A single-amino acid substitution in West Nile virus 2K peptide between NS4A and NS4B confers resistance to lycorine, a flavivirus inhibitor

Zou, Gang; Puig-Basagoiti, Francesc; Zhang, Bo; Qing, Min; Chen, Liqiang; Pankiewicz, Krzysztof W.; Felczak, Krzysztof; Yuan, Zhiming; Shi, Pei Yong

doi:10.1016/j.virol.2008.11.003

Cited by 122 publications

(115 citation statements)

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“…5 and 6D). The same 2K mutation recently was reported to confer WNV resistance to lycorine, an inhibitor of flavivirus RNA synthesis (46). These results suggest that the 2K mutation modulates the ER membrane rearrangement, leading to a better scaffold for replication complex formation.…”

Section: Superinfection Of Wnv Neorep Bhk-21 Cells Using Flavivirusessupporting

confidence: 72%

Exclusion of West Nile Virus Superinfection through RNA Replication

Zou

Zhang

Lim

et al. 2009

J Virol

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Section: Superinfection Of Wnv Neorep Bhk-21 Cells Using Flavivirusessupporting

confidence: 72%

Exclusion of West Nile Virus Superinfection through RNA Replication

Zou

Zhang

Lim

et al. 2009

J Virol

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“…For the luciferase assays, 10 l of lysate was mixed with 50 l of the Renilla luciferase assay reagent (Promega). For quantification of compound-mediated inhibition, the relative luciferase activity derived from the mock-treated cells was set as 100% (78).…”

Section: Methodsmentioning

confidence: 99%

A Novel Dengue Virus Inhibitor, BP13944, Discovered by High-Throughput Screening with Dengue Virus Replicon Cells Selects for Resistance in the Viral NS2B/NS3 Protease

Yang

et al. 2014

Antimicrob Agents Chemother

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bDengue virus (DENV) causes disease globally, resulting in an estimated 25 to 100 million new infections per year. No effective DENV vaccine is available, and the current treatment is only supportive. Thus, there is an urgent need to develop therapeutic agents to cure this epidemic disease. In the present study, we identified a potential small-molecule inhibitor, BP13944, via highthroughput screening (HTS) of 60,000 compounds using a stable cell line harboring an efficient luciferase replicon of DENV serotype 2 (DENV-2). BP13944 reduced the expression of the DENV replicon reporter in cells, showing a 50% effective concentration (EC 50 ) of 1.03 ؎ 0.09 M. Without detectable cytotoxicity, the compound inhibited replication or viral RNA synthesis in all four serotypes of DENV but not in Japanese encephalitis virus (JEV). Sequencing analyses of several individual clones derived from BP13944-resistant RNAs purified from cells harboring the DENV-2 replicon revealed a consensus amino acid substitution (E66G) in the region of the NS3 protease domain. Introduction of E66G into the DENV replicon, an infectious DENV cDNA clone, and recombinant NS2B/NS3 protease constructs conferred 15.2-, 17.2-, and 3.1-fold resistance to BP13944, respectively. Our results identify an effective small-molecule inhibitor, BP13944, which likely targets the DENV NS3 protease. BP13944 could be considered part of a more effective treatment regime for inhibiting DENV in the future. D engue virus (DENV) (serotypes 1 to 4) belongs to the familyFlaviviridae, a group of enveloped RNA viruses that includes the genera Hepacivirus, Flavivirus, and Pestivirus. The genus Flavivirus consists of arthropod-borne disease agents such as yellow fever virus (YFV), Japanese encephalitis virus (JEV), West Nile virus (WNV), tick-borne encephalitis virus (TBEV), and DENV (1). More than 70 members of the Flavivirus genus are important human pathogens that cause significant morbidity and mortality (2). DENV is a public health threat to an estimated 2.5 billion people living in areas where dengue is epidemic, leading to 50 to 100 million human infections each year (3, 4). DENV infection frequently leads to dengue fever, life-threatening dengue hemorrhagic fever (DHF), or dengue shock syndrome (DSS) (5-7). Approximately 500,000 cases of DHF and DSS have been reported among more than 100 countries, causing approximately 12,500 deaths per year (3). Despite the tremendous efforts invested in anti-DENV research, no clinically approved vaccine or antiviral therapeutic agents are available for humans, and disease treatment is limited to supportive care (4,8,9). Considering the spread of this epidemic and the severity of DENV, an effective anti-DENV drug is urgently needed.DENV is an enveloped RNA virus consisting of a 10.7-kb single-stranded, positive-polarity RNA genome associated with multiple copies of capsid proteins. DENV RNA is translated as a single polyprotein upon entering the host cell and is cleaved by host proteases and the virus-encoded two-component protease (NS...

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“…The cells were lysed for luminometry, and a luciferase assay was performed by mixing 5 l of lysate with 25 l of Renilla Luciferase Assay Reagent (Promega). For quantification of the compound-mediated inhibition, the relative luciferase activity derived from the mock-treated cells was set to 100% (48).…”

Section: Methodsmentioning

confidence: 99%

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

Lin

Wang

et al. 2013

Antimicrob Agents Chemother

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bHepatitis C virus (HCV), a member of the Flaviviridae family, 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 (EC 50 ) and a selective index value of 3.9 ؎ 0.9 pM and >12,800,000, respectively. DBPR110 reduced HCV2a replicon activity with an EC 50 and 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-␣), 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. Hepatitis C virus (HCV) is a small enveloped RNA virus that affects nearly 170 million individuals worldwide, making it a leading cause of hepatitis C and liver disease (1). HCV infection is responsible for the development of severe chronic liver disease and cirrhosis and associated complications, including liver failure, portal hypertension, and hepatocellular carcinoma (2). The main goals of chronic HCV therapy are to eradicate the virus and prevent these potentially life-threatening complications. The mainstays of chronic HCV therapy are PEGylated alpha interferon (IFN-␣) and ribavirin, but these compounds are poorly tolerated and may eventually lead to a suboptimal response rate and a high incidence of adverse effects, including flu-like symptoms, depression, and anemia (3, 4). The chances of sustained viral clearance are only 40 to 50% for genotype 1 infection, which is the predominant genotype in worldwide populations. Therefore, the development of specific antiviral therapies for hepatitis C with improved efficacy and better tolerance is a major public health objective that is urgently important.HCV is a positive-strand RNA virus that has been classified as the sole member of the Hepacivirus genus within the Flaviviridae family. The HCV genome consists of a single strand of RNA that is approximately 9.6 kb long, with a large open reading frame encoding a polyprotein of ...

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A single-amino acid substitution in West Nile virus 2K peptide between NS4A and NS4B confers resistance to lycorine, a flavivirus inhibitor

Cited by 122 publications

References 46 publications

Exclusion of West Nile Virus Superinfection through RNA Replication

Exclusion of West Nile Virus Superinfection through RNA Replication

A Novel Dengue Virus Inhibitor, BP13944, Discovered by High-Throughput Screening with Dengue Virus Replicon Cells Selects for Resistance in the Viral NS2B/NS3 Protease

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

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