Cargando…

Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study

The synthetic benzimidazole opioid etazene (which has a 70-times higher analgesic activity than morphine), a recreational drug, has gained popularity as a novel psychoactive substance (NPS) on the illegal/darknet market; however, no experimental information is available at the molecular level on the...

Descripción completa

Detalles Bibliográficos
Autores principales: Chaturvedi, Krishna, Hewamanna, Isuru, Pandey, Pankaj, Khan, Washim, Wang, Yan-Hong, Chittiboyina, Amar G., Doerksen, Robert J., Godfrey, Murrell
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9959024/
https://www.ncbi.nlm.nih.gov/pubmed/36838590
http://dx.doi.org/10.3390/molecules28041601
_version_ 1784895170410446848
author Chaturvedi, Krishna
Hewamanna, Isuru
Pandey, Pankaj
Khan, Washim
Wang, Yan-Hong
Chittiboyina, Amar G.
Doerksen, Robert J.
Godfrey, Murrell
author_facet Chaturvedi, Krishna
Hewamanna, Isuru
Pandey, Pankaj
Khan, Washim
Wang, Yan-Hong
Chittiboyina, Amar G.
Doerksen, Robert J.
Godfrey, Murrell
author_sort Chaturvedi, Krishna
collection PubMed
description The synthetic benzimidazole opioid etazene (which has a 70-times higher analgesic activity than morphine), a recreational drug, has gained popularity as a novel psychoactive substance (NPS) on the illegal/darknet market; however, no experimental information is available at the molecular level on the binding mechanism and putative binding site of etazene and its metabolites at the µ-opioid receptor (MOR). In the present study, we investigated the metabolism of etazene in human liver microsomes using ultra-high-performance liquid chromatography–mass spectrometry (UHPLC–MS). We also explored the possibilities of MOR activation by etazene and its metabolites by studying their binding mechanisms and interaction profiles at an active-state MOR model via molecular docking, binding free energy calculations, and all-atom molecular dynamics (MD) simulations. The putative metabolites of etazene were also predicted using the ADMET Predictor 10.1. The molecular docking studies and free energy calculations showed that etazene and its metabolites (M1, M2, and M5–M7) exhibited strong predicted binding affinity at MOR and showed overlapped binding orientation with MOR-bound agonist BU72, which was co-crystallized in the MOR X-ray crystal structure (PDB ID: 5C1M). MD also confirmed the stability of the MOR–etazene and MOR–M6 complexes. These results suggest that etazene and its metabolites may act as strong MOR agonists, highlighting the necessity of experimental validation. The insights from this study, such as key interactions between etazene and its metabolites and the MOR, will allow authorities to predict potential analogs and clarify the target–protein interactions associated with this illicit substance, granting advanced or rapid reactions to confiscating or banning potential emerging drugs.
format Online
Article
Text
id pubmed-9959024
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-99590242023-02-26 Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study Chaturvedi, Krishna Hewamanna, Isuru Pandey, Pankaj Khan, Washim Wang, Yan-Hong Chittiboyina, Amar G. Doerksen, Robert J. Godfrey, Murrell Molecules Article The synthetic benzimidazole opioid etazene (which has a 70-times higher analgesic activity than morphine), a recreational drug, has gained popularity as a novel psychoactive substance (NPS) on the illegal/darknet market; however, no experimental information is available at the molecular level on the binding mechanism and putative binding site of etazene and its metabolites at the µ-opioid receptor (MOR). In the present study, we investigated the metabolism of etazene in human liver microsomes using ultra-high-performance liquid chromatography–mass spectrometry (UHPLC–MS). We also explored the possibilities of MOR activation by etazene and its metabolites by studying their binding mechanisms and interaction profiles at an active-state MOR model via molecular docking, binding free energy calculations, and all-atom molecular dynamics (MD) simulations. The putative metabolites of etazene were also predicted using the ADMET Predictor 10.1. The molecular docking studies and free energy calculations showed that etazene and its metabolites (M1, M2, and M5–M7) exhibited strong predicted binding affinity at MOR and showed overlapped binding orientation with MOR-bound agonist BU72, which was co-crystallized in the MOR X-ray crystal structure (PDB ID: 5C1M). MD also confirmed the stability of the MOR–etazene and MOR–M6 complexes. These results suggest that etazene and its metabolites may act as strong MOR agonists, highlighting the necessity of experimental validation. The insights from this study, such as key interactions between etazene and its metabolites and the MOR, will allow authorities to predict potential analogs and clarify the target–protein interactions associated with this illicit substance, granting advanced or rapid reactions to confiscating or banning potential emerging drugs. MDPI 2023-02-07 /pmc/articles/PMC9959024/ /pubmed/36838590 http://dx.doi.org/10.3390/molecules28041601 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Chaturvedi, Krishna
Hewamanna, Isuru
Pandey, Pankaj
Khan, Washim
Wang, Yan-Hong
Chittiboyina, Amar G.
Doerksen, Robert J.
Godfrey, Murrell
Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study
title Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study
title_full Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study
title_fullStr Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study
title_full_unstemmed Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study
title_short Identification of the Putative Binding Site of a Benzimidazole Opioid (Etazene) and Its Metabolites at µ-Opioid Receptor: A Human Liver Microsomal Assay and Systematic Computational Study
title_sort identification of the putative binding site of a benzimidazole opioid (etazene) and its metabolites at µ-opioid receptor: a human liver microsomal assay and systematic computational study
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9959024/
https://www.ncbi.nlm.nih.gov/pubmed/36838590
http://dx.doi.org/10.3390/molecules28041601
work_keys_str_mv AT chaturvedikrishna identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy
AT hewamannaisuru identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy
AT pandeypankaj identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy
AT khanwashim identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy
AT wangyanhong identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy
AT chittiboyinaamarg identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy
AT doerksenrobertj identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy
AT godfreymurrell identificationoftheputativebindingsiteofabenzimidazoleopioidetazeneanditsmetabolitesatμopioidreceptorahumanlivermicrosomalassayandsystematiccomputationalstudy