Cargando…

Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity

Two promising lead structures of small molecular orexin receptor agonist have been reported, but without detailed analyses of the pharmacological properties. One of them, 1-(3,4-dichlorophenyl)-2-[2-imino-3-(4-methylbenzyl)-2,3-dihydro-1H-benzo[d]imidazol-1-yl]ethan-1-ol (Yan 7874), is commercially...

Descripción completa

Detalles Bibliográficos
Autores principales: Turku, Ainoleena, Rinne, Maiju K., Boije af Gennäs, Gustav, Xhaard, Henri, Lindholm, Dan, Kukkonen, Jyrki P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5456073/
https://www.ncbi.nlm.nih.gov/pubmed/28575023
http://dx.doi.org/10.1371/journal.pone.0178526
_version_ 1783241166836203520
author Turku, Ainoleena
Rinne, Maiju K.
Boije af Gennäs, Gustav
Xhaard, Henri
Lindholm, Dan
Kukkonen, Jyrki P.
author_facet Turku, Ainoleena
Rinne, Maiju K.
Boije af Gennäs, Gustav
Xhaard, Henri
Lindholm, Dan
Kukkonen, Jyrki P.
author_sort Turku, Ainoleena
collection PubMed
description Two promising lead structures of small molecular orexin receptor agonist have been reported, but without detailed analyses of the pharmacological properties. One of them, 1-(3,4-dichlorophenyl)-2-[2-imino-3-(4-methylbenzyl)-2,3-dihydro-1H-benzo[d]imidazol-1-yl]ethan-1-ol (Yan 7874), is commercially available, and we set out to analyze its properties. As test system we utilized human OX(1) and OX(2) orexin receptor-expressing Chinese hamster ovary (CHO) K1 cells as well as control CHO-K1 and neuro-2a neuroblastoma cells. G(q)-coupling was assessed by measurement of intracellular Ca(2+) and phospholipase C activity, and the coupling to G(i) and G(s) by adenylyl cyclase inhibition and stimulation, respectively. At concentrations above 1 μM, strong Ca(2+) and low phospholipase C responses to Yan 7874 were observed in both OX(1)- and OX(2)-expressing cells. However, a major fraction of the response was not mediated by orexin receptors, as determined utilizing the non-selective orexin receptor antagonist N-biphenyl-2-yl-1-{[(1-methyl-1H-benzimidazol-2-yl)sulfanyl]acetyl}-L-prolinamide (TCS 1102) as well as control CHO-K1 cells. Yan 7874 did not produce any specific adenylyl cyclase response. Some experiments suggested an effect on cell viability by Yan 7874, and we thus analyzed this. Within a few hours of exposure, Yan 7874 markedly changed cell morphology (shrunken, rich in vacuoles), reduced growth, promoted cell detachment, and induced necrotic cell death. The effect was equal in cells expressing orexin receptors or not. Thus, Yan 7874 is a weak partial agonist of orexin receptors. It also displays strong off-target effects in the same concentration range, culminating in necrotic cell demise. This makes Yan 7874 unsuitable as orexin receptor agonist.
format Online
Article
Text
id pubmed-5456073
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-54560732017-06-12 Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity Turku, Ainoleena Rinne, Maiju K. Boije af Gennäs, Gustav Xhaard, Henri Lindholm, Dan Kukkonen, Jyrki P. PLoS One Research Article Two promising lead structures of small molecular orexin receptor agonist have been reported, but without detailed analyses of the pharmacological properties. One of them, 1-(3,4-dichlorophenyl)-2-[2-imino-3-(4-methylbenzyl)-2,3-dihydro-1H-benzo[d]imidazol-1-yl]ethan-1-ol (Yan 7874), is commercially available, and we set out to analyze its properties. As test system we utilized human OX(1) and OX(2) orexin receptor-expressing Chinese hamster ovary (CHO) K1 cells as well as control CHO-K1 and neuro-2a neuroblastoma cells. G(q)-coupling was assessed by measurement of intracellular Ca(2+) and phospholipase C activity, and the coupling to G(i) and G(s) by adenylyl cyclase inhibition and stimulation, respectively. At concentrations above 1 μM, strong Ca(2+) and low phospholipase C responses to Yan 7874 were observed in both OX(1)- and OX(2)-expressing cells. However, a major fraction of the response was not mediated by orexin receptors, as determined utilizing the non-selective orexin receptor antagonist N-biphenyl-2-yl-1-{[(1-methyl-1H-benzimidazol-2-yl)sulfanyl]acetyl}-L-prolinamide (TCS 1102) as well as control CHO-K1 cells. Yan 7874 did not produce any specific adenylyl cyclase response. Some experiments suggested an effect on cell viability by Yan 7874, and we thus analyzed this. Within a few hours of exposure, Yan 7874 markedly changed cell morphology (shrunken, rich in vacuoles), reduced growth, promoted cell detachment, and induced necrotic cell death. The effect was equal in cells expressing orexin receptors or not. Thus, Yan 7874 is a weak partial agonist of orexin receptors. It also displays strong off-target effects in the same concentration range, culminating in necrotic cell demise. This makes Yan 7874 unsuitable as orexin receptor agonist. Public Library of Science 2017-06-02 /pmc/articles/PMC5456073/ /pubmed/28575023 http://dx.doi.org/10.1371/journal.pone.0178526 Text en © 2017 Turku 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Turku, Ainoleena
Rinne, Maiju K.
Boije af Gennäs, Gustav
Xhaard, Henri
Lindholm, Dan
Kukkonen, Jyrki P.
Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity
title Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity
title_full Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity
title_fullStr Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity
title_full_unstemmed Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity
title_short Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity
title_sort orexin receptor agonist yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5456073/
https://www.ncbi.nlm.nih.gov/pubmed/28575023
http://dx.doi.org/10.1371/journal.pone.0178526
work_keys_str_mv AT turkuainoleena orexinreceptoragonistyan7874isaweakagonistoforexinhypocretinreceptorsandshowsorexinreceptorindependentcytotoxicity
AT rinnemaijuk orexinreceptoragonistyan7874isaweakagonistoforexinhypocretinreceptorsandshowsorexinreceptorindependentcytotoxicity
AT boijeafgennasgustav orexinreceptoragonistyan7874isaweakagonistoforexinhypocretinreceptorsandshowsorexinreceptorindependentcytotoxicity
AT xhaardhenri orexinreceptoragonistyan7874isaweakagonistoforexinhypocretinreceptorsandshowsorexinreceptorindependentcytotoxicity
AT lindholmdan orexinreceptoragonistyan7874isaweakagonistoforexinhypocretinreceptorsandshowsorexinreceptorindependentcytotoxicity
AT kukkonenjyrkip orexinreceptoragonistyan7874isaweakagonistoforexinhypocretinreceptorsandshowsorexinreceptorindependentcytotoxicity