qHTS Assay for Inhibitors of Hepatitis C Virus (HCV)
Hepatitis C virus (HCV) infects about 200 million people in the world. Many infected people progress to chronic liver disease including cirrhosis with a risk of developing liver cancer. To date, there is no effective vaccine for hepatitis C. Current therapy based on interferon is only effective in about half of the patients and is associated with significant adverse effects. The fraction of more ..
BioActive Compounds: 11624
Depositor Specified Assays
Hepatitis C virus (HCV) infects about 200 million people in the world. Many infected people progress to chronic liver disease including cirrhosis with a risk of developing liver cancer. To date, there is no effective vaccine for hepatitis C. Current therapy based on interferon is only effective in about half of the patients and is associated with significant adverse effects. The fraction of people with HCV who can complete a successful treatment is estimated to be no more than 10 percent. Recent development of direct-acting antivirals against HCV, such as protease and polymerase inhibitors, is promising but still requires combination with peginterferon and ribavirin for maximal efficacy. In addition, these agents are associated with high rate of resistance and many have significant side effects.
Due to the lack of a culture system for infectious HCV, the search for new HCV drugs has been greatly hampered. Cell-based screen for HCV inhibitors in use today is based on the HCV replicon system, which only targets the RNA replication step of the viral lifecycle and does not encompass viral entry, processing, assembly and secretion. High-throughput screening (HTS) with an infectious HCV system would cover the complete spectrum of potentially druggable targets in all stages of HCV lifecycle, and would have more biological relevance than other cell-based assays. Moreover, targeting several key processes in the viral life cycle may not only increase antiviral efficacy; more importantly, it may also reduce the capacity of the virus to develop resistance to the compound.
The goal of this project is to identify novel HCV inhibitors as new therapies for hepatitis C, using a highly sensitive and specific assay platform which is based on a HCV infectious cell culture system established in the laboratory and adapted for high-throughput HCV drug screen.
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]
MLPCN Grant: MH095511
Assay Submitter (PI): Jake Liang, NIDDK
The assay will start with plating 1,000 cells/well in 3 muL volume and culture for 4 h. Then 23 nL of compounds from the library collection will be added to each well, followed by adding 2.5 muL of HCVcc-Cre virus (~ 0.5 moi) and further cultured for 44 h before the luciferase assay. A volume of 4.5 muL luciferase substrates will be added to each well and the plates will be incubated at room temperature for 15 min. and then read for 15 sec. for the luciferase activity
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
* Activity Concentration. ** Test Concentration.
Data Table (Concise)