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Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors

HIV-1 maturation inhibitors (MIs) disrupt the final step in the HIV-1 protease-mediated cleavage of the Gag polyprotein between capsid p24 capsid (CA) and spacer peptide 1 (SP1), leading to the production of infectious virus. BMS-955176 is a second generation MI with improved antiviral activity towa...

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Autores principales: Lin, Zeyu, Cantone, Joseph, Lu, Hao, Nowicka-Sans, Beata, Protack, Tricia, Yuan, Tian, Yang, Hong, Liu, Zheng, Drexler, Dieter, Regueiro-Ren, Alicia, Meanwell, Nicholas A., Cockett, Mark, Krystal, Mark, Lataillade, Max, Dicker, Ira B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5125710/
https://www.ncbi.nlm.nih.gov/pubmed/27893830
http://dx.doi.org/10.1371/journal.ppat.1005990
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author Lin, Zeyu
Cantone, Joseph
Lu, Hao
Nowicka-Sans, Beata
Protack, Tricia
Yuan, Tian
Yang, Hong
Liu, Zheng
Drexler, Dieter
Regueiro-Ren, Alicia
Meanwell, Nicholas A.
Cockett, Mark
Krystal, Mark
Lataillade, Max
Dicker, Ira B.
author_facet Lin, Zeyu
Cantone, Joseph
Lu, Hao
Nowicka-Sans, Beata
Protack, Tricia
Yuan, Tian
Yang, Hong
Liu, Zheng
Drexler, Dieter
Regueiro-Ren, Alicia
Meanwell, Nicholas A.
Cockett, Mark
Krystal, Mark
Lataillade, Max
Dicker, Ira B.
author_sort Lin, Zeyu
collection PubMed
description HIV-1 maturation inhibitors (MIs) disrupt the final step in the HIV-1 protease-mediated cleavage of the Gag polyprotein between capsid p24 capsid (CA) and spacer peptide 1 (SP1), leading to the production of infectious virus. BMS-955176 is a second generation MI with improved antiviral activity toward polymorphic Gag variants compared to a first generation MI bevirimat (BVM). The underlying mechanistic reasons for the differences in polymorphic coverage were studied using antiviral assays, an LC/MS assay that quantitatively characterizes CA/SP1 cleavage kinetics of virus like particles (VLPs) and a radiolabel binding assay to determine VLP/MI affinities and dissociation kinetics. Antiviral assay data indicates that BVM does not achieve 100% inhibition of certain polymorphs, even at saturating concentrations. This results in the breakthrough of infectious virus (partial antagonism) regardless of BVM concentration. Reduced maximal percent inhibition (MPI) values for BVM correlated with elevated EC(50) values, while rates of HIV-1 protease cleavage at CA/SP1 correlated inversely with the ability of BVM to inhibit HIV-1 Gag polymorphic viruses: genotypes with more rapid CA/SP1 cleavage kinetics were less sensitive to BVM. In vitro inhibition of wild type VLP CA/SP1 cleavage by BVM was not maintained at longer cleavage times. BMS-955176 exhibited greatly improved MPI against polymorphic Gag viruses, binds to Gag polymorphs with higher affinity/longer dissociation half-lives and exhibits greater time-independent inhibition of CA/SP1 cleavage compared to BVM. Virological (MPI) and biochemical (CA/SP1 cleavage rates, MI-specific Gag affinities) data were used to create an integrated semi-quantitative model that quantifies CA/SP1 cleavage rates as a function of both MI and Gag polymorph. The model outputs are in accord with in vitro antiviral observations and correlate with observed in vivo MI efficacies. Overall, these findings may be useful to further understand antiviral profiles and clinical responses of MIs at a basic level, potentially facilitating further improvements to MI potency and coverage.
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spelling pubmed-51257102016-12-15 Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors Lin, Zeyu Cantone, Joseph Lu, Hao Nowicka-Sans, Beata Protack, Tricia Yuan, Tian Yang, Hong Liu, Zheng Drexler, Dieter Regueiro-Ren, Alicia Meanwell, Nicholas A. Cockett, Mark Krystal, Mark Lataillade, Max Dicker, Ira B. PLoS Pathog Research Article HIV-1 maturation inhibitors (MIs) disrupt the final step in the HIV-1 protease-mediated cleavage of the Gag polyprotein between capsid p24 capsid (CA) and spacer peptide 1 (SP1), leading to the production of infectious virus. BMS-955176 is a second generation MI with improved antiviral activity toward polymorphic Gag variants compared to a first generation MI bevirimat (BVM). The underlying mechanistic reasons for the differences in polymorphic coverage were studied using antiviral assays, an LC/MS assay that quantitatively characterizes CA/SP1 cleavage kinetics of virus like particles (VLPs) and a radiolabel binding assay to determine VLP/MI affinities and dissociation kinetics. Antiviral assay data indicates that BVM does not achieve 100% inhibition of certain polymorphs, even at saturating concentrations. This results in the breakthrough of infectious virus (partial antagonism) regardless of BVM concentration. Reduced maximal percent inhibition (MPI) values for BVM correlated with elevated EC(50) values, while rates of HIV-1 protease cleavage at CA/SP1 correlated inversely with the ability of BVM to inhibit HIV-1 Gag polymorphic viruses: genotypes with more rapid CA/SP1 cleavage kinetics were less sensitive to BVM. In vitro inhibition of wild type VLP CA/SP1 cleavage by BVM was not maintained at longer cleavage times. BMS-955176 exhibited greatly improved MPI against polymorphic Gag viruses, binds to Gag polymorphs with higher affinity/longer dissociation half-lives and exhibits greater time-independent inhibition of CA/SP1 cleavage compared to BVM. Virological (MPI) and biochemical (CA/SP1 cleavage rates, MI-specific Gag affinities) data were used to create an integrated semi-quantitative model that quantifies CA/SP1 cleavage rates as a function of both MI and Gag polymorph. The model outputs are in accord with in vitro antiviral observations and correlate with observed in vivo MI efficacies. Overall, these findings may be useful to further understand antiviral profiles and clinical responses of MIs at a basic level, potentially facilitating further improvements to MI potency and coverage. Public Library of Science 2016-11-28 /pmc/articles/PMC5125710/ /pubmed/27893830 http://dx.doi.org/10.1371/journal.ppat.1005990 Text en © 2016 Lin et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Lin, Zeyu
Cantone, Joseph
Lu, Hao
Nowicka-Sans, Beata
Protack, Tricia
Yuan, Tian
Yang, Hong
Liu, Zheng
Drexler, Dieter
Regueiro-Ren, Alicia
Meanwell, Nicholas A.
Cockett, Mark
Krystal, Mark
Lataillade, Max
Dicker, Ira B.
Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors
title Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors
title_full Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors
title_fullStr Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors
title_full_unstemmed Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors
title_short Mechanistic Studies and Modeling Reveal the Origin of Differential Inhibition of Gag Polymorphic Viruses by HIV-1 Maturation Inhibitors
title_sort mechanistic studies and modeling reveal the origin of differential inhibition of gag polymorphic viruses by hiv-1 maturation inhibitors
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5125710/
https://www.ncbi.nlm.nih.gov/pubmed/27893830
http://dx.doi.org/10.1371/journal.ppat.1005990
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