Author:

Keywords:

Science & Technology, Life Sciences & Biomedicine, Chemistry, Medicinal, Pharmacology & Pharmacy, ALPHA-AMINO-ACIDS, LEDGF/P75 PROTEIN, P53 PEPTIDE, SIDE-CHAIN, BH3 HELIX, IN-VIVO, DOLUTEGRAVIR, REPLICATION, DISCOVERY, INFECTION, Adaptor Proteins, Signal Transducing, Amino Acid Sequence, Biocatalysis, Cell Line, Tumor, Cell Membrane Permeability, Cell Survival, Drug Design, Enzyme Inhibitors, HCT116 Cells, HIV Integrase, HIV-1, Host-Pathogen Interactions, Humans, Microscopy, Confocal, Models, Chemical, Molecular Structure, Peptides, Precursor Cell Lymphoblastic Leukemia-Lymphoma, Protein Binding, Transcription Factors, Virus Replication, 0304 Medicinal and Biomolecular Chemistry, 0305 Organic Chemistry, 1115 Pharmacology and Pharmaceutical Sciences, Medicinal & Biomolecular Chemistry, 3214 Pharmacology and pharmaceutical sciences, 3404 Medicinal and biomolecular chemistry, 3405 Organic chemistry

Abstract:

DESIGN OF CELL-PERMEABLE NANONEEDLES AS HIV-1 INTEGRASE INHIBITORS Ya-Qiu Long*,†, Shao-Xu Huang†, ‡, Zahrah Zawahir§,‡, Huiyuan Li†, Tino W. Sanchez§, Ying Zhi†, Frauke Christ¶, Zeger Debyser¶, Nouri Neamati*,§ †State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China; §Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, Los Angeles, CA 90033; ¶The Laboratory for Molecular Virology and Gene Therapy, KULeuven and IRC KULAK, Kapucijnenvoer 33, B-3000 Leuven, Flanders, Belgium HIV-1 integrase (IN) is an essential enzyme in the viral life cycle and catalyzes the integration of viral DNA into the host genome. It is an attractive therapeutic target for developing HIV-specific drugs because no mammalian counterpart exists for this enzyme. For successful integration of reverse-transcribed viral DNA into the host genome to occur, several interactions with host proteins are required. We have previously demonstrated HIV-1 IN inhibition in vitro with peptides derived from naturally occurring alpha-helical regions of the protein. Two peptides, NL6 and NL9, with low micromolar IC50 values for inhibition of HIV-1 IN catalysis were discovered. Each peptide was derived from the α1 and α3 helical domains of the IN protein, respectively. Alanine scanning on these peptides further pinpointed amino acid residues that were later proven to be critical for IN dimerization and LEDGF/p75-IN interaction. Here we show that hydrocarbon-stapling of these peptides to stabilize their helical structure enables enhanced enzyme inhibitory potency and cell permeability while demonstrating selective inhibition of HIV-1 replication in cell culture. Furthermore, the stabilized peptides demonstrate inhibition of the LEDGF/p75 interaction. The corresponding unstapled peptides do not show inhibition of replication in vivo, although each pair of peptides has similar activity against IN in our in vitro assay. Given their enhanced potency and cell permeability they may serve as prototypical biochemical probes for development into ‘nanoneedles’ for the elucidation of HIV-1 IN and host co-factor interactions within their native cellular environment. To our knowledge this is the first report attempting to design and synthesize stapled-peptides targeting integration events either by directly inhibiting IN or inhibiting its interaction with LEDGF/p75.