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Genotype 1b (Con1, Licensed from Reblikon GmbH) HCV Replicon Cells Were Resuspended to a Concentration of 1.4 × 105 Cells/ml by Addition of Pre-Warmed Medium (DMEM + 10% FCS). 45 μl of This Suspension Was Added to Each Well a 384-Well Assay Plate (Lumitrac, Greiner) Already Containing 0.5 l of Test Compound. All Plates Were Covered with gas Permeable Seals and Incubated at 37oC, 5% CO2 for 48 Hours. After 48 Hours, the Plate Was Removed from the Incubator and Left to Cool to Room Temperature for 15 -30 mins. An Equal Volume of Reconstituted Lyophilised Britelite plus Substrate (PerkinElmer) to Medium Was Added to Each Well. Luminescence Was Read Immediately on an EnVision (Perkin Elmer) Plate Reader.

has been cited by the following article:

• TITLE: Identification of HCV Inhibitors from a Cell-Based Sub-Genomic Replicon Screen

  AUTHORS: David C. Pryde, Thien-Duc Tran, Mark Gardner, Chris Pickford, Stephen M. Shaw, Mike Westby, Tanya Parkinson, Caroline Smith-Burchnell, Rob Webster, Satish Dayal

  KEYWORDS: HCV; Replicon; Antiviral; Cell-Based Screen

  JOURNAL NAME: Open Journal of Medicinal Chemistry, Vol.3 No.1, March 29, 2013

  ABSTRACT: A high throughput screen of the Pfizer compound collection was carried out using a hepatitis C virus (HCV) genotype 1b subgenomic replicon cell line. Those confirmed hits that demonstrated broad spectrum activity without overt cytotoxicity were further evaluated, leading to the identification of a series of pyrrolopyridines with excellent antiviral activity in a fully infectious HCV cell-based assay and pharmacokinetic properties.