Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Apr 26;62(5):e02060-17.
doi: 10.1128/AAC.02060-17. Print 2018 May.

GRL-079, a Novel HIV-1 Protease Inhibitor, Is Extremely Potent against Multidrug-Resistant HIV-1 Variants and Has a High Genetic Barrier against the Emergence of Resistant Variants

Nicole S Delino  1   2 , Manabu Aoki  1 , Hironori Hayashi  2 , Shin-Ichiro Hattori  2 , Simon B Chang  1 , Yuki Takamatsu  1 , Cuthbert D Martyr  3   4 , Debananda Das  1 , Arun K Ghosh  3   4 , Hiroaki Mitsuya  5   2   6   7
Affiliations

GRL-079, a Novel HIV-1 Protease Inhibitor, Is Extremely Potent against Multidrug-Resistant HIV-1 Variants and Has a High Genetic Barrier against the Emergence of Resistant Variants

Nicole S Delino et al. Antimicrob Agents Chemother. .

Abstract

We identified four novel nonpeptidic human immunodeficiency virus type 1 (HIV-1) protease inhibitors (PIs), GRL-078, -079, -077, and -058, containing an alkylamine at the C-5 position of P2 tetrahydropyrano-tetrahydrofuran (Tp-THF) and a P2' cyclopropyl (Cp) (or isopropyl)-aminobenzothiazole (Abt) moiety. Their 50% effective concentrations (EC50s) were 2.5 to 30 nM against wild-type HIV-1NL4-3, 0.3 to 6.7 nM against HIV-2EHO, and 0.9 to 90 nM against laboratory-selected PI-resistant HIV-1 and clinical HIV-1 variants resistant to multiple FDA-approved PIs (HIVMDR). GRL-078, -079, -077, and -058 also effectively blocked the replication of HIV-1 variants highly resistant to darunavir (DRV) (HIVDRVrp51), with EC50s of 38, 62, 61, and 90 nM, respectively, while four FDA-approved PIs examined (amprenavir, atazanavir, lopinavir [LPV], and DRV) had virtually no activity (EC50s of >1,000 nM) against HIVDRVrp51 Structurally, GRL-078, -079, and -058 form strong hydrogen bond interactions between Tp-THF modified at C-5 and Asp29/Asp30/Gly48 of wild-type protease, while the P2' Cp-Abt group forms strong hydrogen bonds with Asp30'. The Tp-THF and Cp-Abt moieties also have good nonpolar interactions with protease residues located in the flap region. For selection with LPV and DRV by use of a mixture of 11 HIVMDR strains (HIV11MIX), HIV11MIX became highly resistant to LPV and DRV over 13 to 32 and 32 to 41 weeks, respectively. However, for selection with GRL-079 and GRL-058, HIV11MIX failed to replicate at >0.08 μM and >0.2 μM, respectively. Thermal stability results supported the highly favorable anti-HIV-1 potency of GRL-079 as well as other PIs. The present data strongly suggest that the P2 Tp-THF group modified at C-5 and the P2' Abt group contribute to the potent anti-HIV-1 profiles of the four PIs against HIV-1NL4-3 and a wide spectrum of HIVMDR strains.

Keywords: AIDS; HIV-1; protease; protease inhibitors.

PubMed Disclaimer

Figures

FIG 1
FIG 1
(A) Structures of GRL-0476, -015, -038, -078, -079, -077, and -058. (B) The structure of darunavir is shown as a reference.
FIG 2
FIG 2
Structural modeling of PRWT complexed with GRL-078, -079, and -058. Polar contacts with PRWT made by GRL-078 (A), GRL-079 (B), GRL-058 (C), and DRV (D) are shown. One protease subunit is shown in green, while the other, identical subunit is shown in red. In each panel, the carbons for the protease inhibitors are displayed in green, while PRWT carbons are shown in gray. Nitrogen, oxygen, and sulfur atoms are shown in blue, red, and yellow, respectively. Yellow dotted lines indicate hydrogen bonds.
FIG 3
FIG 3
Connolly molecular surface interactions of GRL-078 (A), GRL-079 (B), and DRV (C) with selected protease residues located in the flap region. The inhibitor surfaces are shown in gray, and the surfaces for I50′, I47′, K45′, and G48 are shown in magenta, brown, green, and blue, respectively. The protease chains are shown in green and red. In each panel, the carbons for the protease inhibitors are displayed in green, while PRWT carbons are shown in gray. Nitrogen, oxygen, and sulfur atoms are shown in blue, red, and yellow, respectively.
FIG 4
FIG 4
In vitro selection of HIV-1 variants against GRL-079, GRL-058, DRV, and LPV. A mixture of 11 multi-PI-resistant HIV-1 isolates (HIV11MIX) was propagated in MT-4 cells in the presence of increasing concentrations of GRL-079 (○), GRL-058 (▲), DRV (□), or LPV (●). The selection process was conducted by passage of cell-free virus. Amino acid substitutions acquired in each HIV-1 variant during selection are illustrated in Fig. 5.
FIG 5
FIG 5
Amino acid sequences of the proteases of HIV11MIX variants selected with LPV, DRV, GRL-079, and GRL-058 in vitro. Amino acid sequences deduced from the nucleotide sequences of the protease-encoding region of proviral DNA isolated from HIV11MIX variants selected with LPV (13 weeks for experiment 1 and 32 weeks for experiment 2) (A), DRV (32 weeks for experiment 1 and 41 weeks for experiment 2) (B), GRL-079 (32 weeks for experiment 1 and 37 weeks for experiment 2) (C), and GRL-058 (32 weeks for experiment 1 and 44 weeks for experiment 2) (D) are shown. The HIVNL4-3 and HIV11MIXWK0 amino acid sequences are displayed at the top for reference. Identity at individual amino acid positions is indicated by dots. Fractions shown on the right give the number of viruses from which each clone is presumed to have originated over the total number of clones examined.
FIG 6
FIG 6
Thermal stability of PRD25N with DRV or GRL-0476, -015, -038, -058, -077, -078, or -079 as determined using differential scanning fluorimetry (DSF). (A) Relative fluorescence intensities determined by DSF using Sypro Orange. (B) Tm values shown represent the temperatures at which the relative fluorescence intensity was 0.5, and ΔTm values indicate the Tm difference between protease complexed with each PI and that with no PI. SD, standard deviation.
FIG 7
FIG 7
Amino acid sequences of the proteases of 11 HIVMDR strains. The amino acid sequences of protease deduced from nucleotide sequences of the protease-encoding region for 11 HIVMDR strains are shown. The consensus sequence of HIV-1NL4-3 is illustrated at the top as a reference. Identity at individual amino acid positions is indicated by dots.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Walensky RP, Paltiel AD, Losina E, Mercincavage LM, Schackman BR, Sax PE, Weinstein MC, Freedberg KA. 2006. The survival benefits of AIDS treatment in the United States. J Infect Dis 194:11–19. doi:10.1086/505147. - DOI - PubMed
    1. Edmonds A, Yotebieng M, Lusiama J, Matumona Y, Kitetele F, Napravnik S, Cole SR, Van Rie A, Behets F. 2011. The effect of highly active antiretroviral therapy on the survival of HIV-infected children in a resource-deprived setting: a cohort study. PLoS Med 8:e1001044. doi:10.1371/journal.pmed.1001044. - DOI - PMC - PubMed
    1. Lohse N, Hansen AB, Gerstoft J, Obel N. 2007. Improved survival in HIV-infected persons: consequences and perspectives. J Antimicrob Chemother 60:461–463. doi:10.1093/jac/dkm241. - DOI - PubMed
    1. Gomes MJ, Neves J, Sarmento B. 2014. Nanoparticle-based drug delivery to improve the efficacy of antiretroviral therapy in the central nervous system. Int J Nanomedicine (Lond) 9:1757–1769. doi:10.2147/IJN.S45886. - DOI - PMC - PubMed
    1. Pantaleo G, Graziosi C, Butini L, Pizzo PA, Schnittman SM, Kotler DP, Fauci AS. 1991. Lymphoid organs function as major reservoirs for human immunodeficiency virus. Proc Natl Acad Sci U S A 88:9838–9842. - PMC - PubMed

Publication types

LinkOut - more resources