Phage display-directed discovery of LEDGF/p75 binding cyclic peptide inhibitors of HIV replication

Mol Ther. 2012 Nov;20(11):2064-75. doi: 10.1038/mt.2012.132. Epub 2012 Jul 24.

Abstract

The interaction between the human immunodeficiency virus (HIV) integrase (IN) and its cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is crucial for HIV replication. While recently discovered LEDGINs inhibit HIV-1 replication by occupying the LEDGF/p75 pocket in IN, it remained to be demonstrated whether LEDGF/p75 by itself can be targeted. By phage display we identified cyclic peptides (CPs) as the first LEDGF/p75 ligands that inhibit the LEDGF/p75-IN interaction. The CPs inhibit HIV replication in different cell lines without overt toxicity. In accord with the role of LEDGF/p75 in HIV integration and its inhibition by LEDGINs, CP64, and CP65 block HIV replication primarily by inhibiting the integration step. The CPs retained activity against HIV strains resistant to raltegravir or LEDGINs. Saturation transfer difference (STD) NMR showed residues in CP64 that strongly interact with LEDGF/p75 but not with HIV IN. Mutational analysis identified tryptophan as an important residue responsible for the activity of the peptides. Serial passaging of virus in the presence of CPs did not yield resistant strains. Our work provides proof-of-concept for direct targeting of LEDGF/p75 as novel therapeutic strategy and the CPs thereby serve as scaffold for future development of new HIV therapeutics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / antagonists & inhibitors*
  • Adaptor Proteins, Signal Transducing / chemistry
  • Amino Acid Sequence
  • Anti-HIV Agents / chemistry
  • Anti-HIV Agents / pharmacology*
  • Binding Sites
  • Cell Surface Display Techniques*
  • Conserved Sequence
  • Drug Evaluation, Preclinical
  • Drug Resistance, Viral
  • HIV Integrase / chemistry
  • HIV-1 / drug effects
  • HIV-1 / physiology*
  • HIV-2 / drug effects
  • HIV-2 / physiology
  • HeLa Cells
  • Humans
  • Peptide Library
  • Peptides, Cyclic / chemistry
  • Peptides, Cyclic / genetics
  • Peptides, Cyclic / pharmacology*
  • Protein Binding
  • Transcription Factors / antagonists & inhibitors*
  • Transcription Factors / chemistry
  • Virus Internalization
  • Virus Replication*

Substances

  • Adaptor Proteins, Signal Transducing
  • Anti-HIV Agents
  • PSIP1 protein, human
  • Peptide Library
  • Peptides, Cyclic
  • Transcription Factors
  • HIV Integrase