Abstract
A fragment-based screen against human immunodeficiency virus type 1 (HIV) integrase led to a number of compounds that bound to the lens epithelium derived growth factor (LEDGF) binding site of the integrase catalytic core domain. We determined the crystallographic structures of complexes of the HIV integrase catalytic core domain for 10 of these compounds and quantitated the binding by surface plasmon resonance. We demonstrate that the compounds inhibit the interaction of LEDGF with HIV integrase in a proximity AlphaScreen assay, an assay for the LEDGF enhancement of HIV integrase strand transfer and in a cell based assay. The compounds identified represent a potential framework for the development of a new series of HIV integrase inhibitors that do not bind to the catalytic site of the enzyme.
Highlights
Active antiretroviral therapies (HAART) have been an effective way to slow the spread of human immunodeficiency virus type 1 (HIV), but the virus continues to develop resistance to all drugs that have been used in treatment to date [1]
We performed a fragment screen of 500 compounds using surface plasmon resonance (SPR), NMR and crystallography using the His-tagged IN-catalytic core domain (CCD) protein from E. coli
Based on the hits obtained from this screen, several analogues were chosen from the CSIRO compound library and these were tested via SPR and crystallography for binding affinity and the location of binding respectively
Summary
Active antiretroviral therapies (HAART) have been an effective way to slow the spread of HIV, but the virus continues to develop resistance to all drugs that have been used in treatment to date [1]. The structure of HIV IN has been thoroughly investigated [3,4,5,6,7], and consists of three domains (N-terminal DNA binding, catalytic core (CCD) and Cterminal DNA binding). IN forms a complex with viral DNA and several host cellular factors that has been termed the pre-integration complex (PIC) [13]. One component of this complex is lens epithelium derived growth factor (LEDGF/p75), which has a conserved IN binding domain (IBD, residues 347–429) that mediates binding [14]. An epidemiological study of polymorphisms in the PSIP1 gene that codes for the LEDGF protein has shown variation in serologic levels of HIV virus and pathogenesis, consistent with the role of LEDGF in disease progression [19,20]
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