title: A conserved hydrogen-bonding network of P2 bis-tetrahydrofuran-containing HIV-1 protease inhibitors (PIs) with a protease active-site amino acid backbone aids in their activity against PI-resistant HIV.
abstract: In the present study, GRL008, a novel nonpeptidic human immunodeficiency virus type 1 (HIV-1) protease inhibitor (PI), and darunavir (DRV), both of which contain a P2-bis-tetrahydrofuranyl urethane (bis-THF) moiety, were found to exert potent antiviral activity (50% effective concentrations [EC50s], 0.029 and 0.002 μM, respectively) against a multidrug-resistant clinical isolate of HIV-1 (HIVA02) compared to ritonavir (RTV; EC50, >1.0 μM) and tipranavir (TPV; EC50, 0.364 μM). Additionally, GRL008 showed potent antiviral activity against an HIV-1 variant selected in the presence of DRV over 20 passages (HIVDRV(R)P20), with a 2.6-fold increase in its EC50 (0.097 μM) compared to its corresponding EC50 (0.038 μM) against wild-type HIV-1NL4-3 (HIVWT). Based on X-ray crystallographic analysis, both GRL008 and DRV showed strong hydrogen bonds (H-bonds) with the backbone-amide nitrogen/carbonyl oxygen atoms of conserved active-site amino acids G27, D29, D30, and D30' of HIVA02 protease (PRA02) and wild-type PR in their corresponding crystal structures, while TPV lacked H-bonds with G27 and D30' due to an absence of polar groups. The P2' thiazolyl moiety of RTV showed two conformations in the crystal structure of the PRA02-RTV complex, one of which showed loss of contacts in the S2' binding pocket of PRA02, supporting RTV's compromised antiviral activity (EC50, >1 μM). Thus, the conserved H-bonding network of P2-bis-THF-containing GRL008 with the backbone of G27, D29, D30, and D30' most likely contributes to its persistently greater antiviral activity against HIVWT, HIVA02, and HIVDRV(R)P20.
authors.affiliation: Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Ravikiran S
authors.initials: RS
authors.lastname: Yedidi
authors.affiliation: Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Harisha
authors.initials: H
authors.lastname: Garimella
authors.affiliation: Departments of Hematology and Infectious Diseases, Kumamoto University Graduate School of Biomedical Sciences, Kumamoto, Japan Department of Medical Technology, Kumamoto Health Science University, Kumamoto, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Aoki
authors.affiliation: Departments of Hematology and Infectious Diseases, Kumamoto University Graduate School of Biomedical Sciences, Kumamoto, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Aoki-Ogata
authors.affiliation: Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Darshan V
authors.initials: DV
authors.lastname: Desai
authors.affiliation: Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Simon B
authors.initials: SB
authors.lastname: Chang
authors.affiliation: Retroviral Disease Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Davis
authors.affiliation: Departments of Chemistry and Medicinal Chemistry, Purdue University, West Lafayette, Indiana, USA.
authors.forename: W Sean
authors.initials: WS
authors.lastname: Fyvie
authors.affiliation: Protein Expression Laboratory, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Kaufman
authors.affiliation: Life Sciences Collaborative Access Team, Synchrotron Research Center, Northwestern University, Argonne, Illinois, USA.
authors.forename: David W
authors.initials: DW
authors.lastname: Smith
authors.affiliation: Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Debananda
authors.initials: D
authors.lastname: Das
authors.affiliation: Protein Expression Laboratory, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Paul T
authors.initials: PT
authors.lastname: Wingfield
authors.affiliation: Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Kenji
authors.initials: K
authors.lastname: Maeda
authors.affiliation: Departments of Chemistry and Medicinal Chemistry, Purdue University, West Lafayette, Indiana, USA.
authors.forename: Arun K
authors.initials: AK
authors.lastname: Ghosh
authors.affiliation: Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA Departments of Hematology and Infectious Diseases, Kumamoto University Graduate School of Biomedical Sciences, Kumamoto, Japan mitsuyah@helix.nih.gov.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mitsuya
mesh_terms: D002219:Carbamates / Q000494:pharmacology*; D020134:Catalytic Domain / Q000187:drug effects*; D003460:Crystallization; D000069454:Darunavir; D024921:Drug Resistance, Multiple, Viral / Q000187:drug effects*; D005663:Furans / Q000494:pharmacology*; D016333:HIV Protease; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology*; D006801:Humans; D006860:Hydrogen Bonding; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D017510:Protein Folding; D011725:Pyridines / Q000494:pharmacology; D011753:Pyrones / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology*; D014961:X-Ray Diffraction
chemical_list: D002219:Carbamates; D005663:Furans; C000601131:GRL008; D017320:HIV Protease Inhibitors; D011725:Pyridines; D011753:Pyrones; D013449:Sulfonamides; D016333:HIV Protease; C514803:p16 protease, Human immunodeficiency virus 1; D000069454:Darunavir; C107201:tipranavir
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