Homogeneous High-Throughput Screening Assays for HIV-1 Integrase 3'-Processing and Strand Transfer Activities

HIV-1 integrase (HIV-IN) is a well-validated antiviral drug target catalyzing a multistep reaction to incorporate the HIV-1 provirus into the genome of the host cell. Smallmolecule inhibitors of HIV-1 integrase that specifically target the strand transfer step have demonstrated efficacy in the suppr...

Full description

Saved in:
Bibliographic Details
Published inSLAS discovery Vol. 10; no. 5; pp. 456 - 462
Main Authors Wang, Yu, Klock, Heath, Yin, Hong, Wolff, Karen, Bieza, Kimberly, Niswonger, Kirk, Matzen, Jason, Gunderson, Drew, Hale, Joanna, Lesley, Scott, Kuhen, Kelli, Caldwell, Jeremy, Brinker, Achim
Format Journal Article
LanguageEnglish
Published Elsevier Inc 01.08.2005
Subjects
3' processing
antivirals
HIV-1
strand transfer
TR-FRET
HIV-1
antivirals
strand transfer
3' processing
TR-FRET
Online AccessGet full text

Cover

More Information
Summary:HIV-1 integrase (HIV-IN) is a well-validated antiviral drug target catalyzing a multistep reaction to incorporate the HIV-1 provirus into the genome of the host cell. Smallmolecule inhibitors of HIV-1 integrase that specifically target the strand transfer step have demonstrated efficacy in the suppression of virus propagation. However, only fewspecific strand transfer inhibitors have been identified to date, and the need to screen for novel compound scaffolds persists. Here, the authors describe 2 homogeneous time-resolved fluorescent resonance energy transfer-based assays for the measurement of HIV-1 integrase 3'-processing and strand transfer activities. Both assayswere optimized for high-throughput screening formats, and a diverse library containingmore than 1million compoundswas screened in 1536-well plates for HIV-IN strand transfer inhibitors. As a result, compounds were found that selectively affect the enzymatic strand transfer reaction over 3' processing. Moreover, several bioactivemoleculeswere identified that inhibited HIV-1 reporter virus infection in cellularmodel systems. In conclusion, the assays presented herein have proven their utility for the identification ofmechanistically interesting and biologically active inhibitors of HIV-1 integrase that hold potential for further development into potent antiviral drugs. (Journal of Biomolecular Screening 2005:456-462)
ISSN:2472-5552
2472-5560
DOI:10.1177/1087057105275212