Development and Validation of a High Throughput Screen for Compounds with Antiviral Activity Against Encephalitic Alphaviruses
Abstract
A high throughput assay using a luciferase tagged VEEV has been established to generate effective probes against encephalitic alphaviruses. The virus was rescued from in vitro transcribed RNA in BHK cells and initial validation has been done. The assay was adapted in an HTS automated form with 384-well system after it has been tested in various conditions including cell number, MOI optimization. Z' value which indicates the robustness of an assay was above 0.6 indicating suitable for a single dose HTS. The PrestWick library, 1120 compounds and Arbovirus hit library, 658 compounds have been tested in duplicate mode. In combination with cytotoxicity of the compounds, 63 compounds were screened out as active hits with > 50% efficacy and < 30% cytotoxicity. Currently those 63 hits were further characterized in a dose response manner. A diverse library consisting of 8960 compounds was tested in the system twice. 16 compounds meeting the criteria were selected and tested further in a dose response format for cytotoxicity and efficacy. A total of 39 compounds which have structural similarity of the selected candidates were tested in a CPE based assay using TC-83.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 15, 2010
- Accession Number
- ADA540027
Entities
People
- Colleen B. Jonsson
- Dong H. Chang
Organizations
- Southern Research