Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification

Abstract Quantification of drug‐metabolizing cytochrome P450 (CYP) isoforms using LC–MS/MS has been proposed as a potential way of estimating antemortem CYP levels using postmortem tissue, but the postmortem stability of CYP proteins is incompletely investigated. If one can use data obtained from th...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Kata W. Pedersen, Jakob Hansen, Jørgen B. Hasselstrøm, Jakob R. Jornil
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2021
Materias:	CYP1A2 CYP2D25 CYP2E1 CYP3A29 cytochrome P450 drug metabolism Therapeutics. Pharmacology RM1-950
Acceso en línea:	https://doaj.org/article/582ec299dd6a4b45822c779ccfcbe8c8
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:582ec299dd6a4b45822c779ccfcbe8c8
record_format	dspace
spelling	oai:doaj.org-article:582ec299dd6a4b45822c779ccfcbe8c82021-11-16T13:45:55ZStability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification2052-170710.1002/prp2.860https://doaj.org/article/582ec299dd6a4b45822c779ccfcbe8c82021-10-01T00:00:00Zhttps://doi.org/10.1002/prp2.860https://doaj.org/toc/2052-1707Abstract Quantification of drug‐metabolizing cytochrome P450 (CYP) isoforms using LC–MS/MS has been proposed as a potential way of estimating antemortem CYP levels using postmortem tissue, but the postmortem stability of CYP proteins is incompletely investigated. If one can use data obtained from the analysis of postmortem specimens to inform physiologically based pharmacokinetic (PBPK) models this greatly increases the access to rare specimens among special subpopulations. In this study, we developed and validated an LC–MS/MS method for targeted CYP protein quantification in a porcine animal model to study postmortem stability. We measured 19.9–28.3 pmol CYP1A2, 50.3–66.2 pmol CYP2D25, 132.9–142.7 pmol CYP2E1, and 16.8–48 pmol CYP3A29 protein per mg PLM in nondegraded tissue. In tissue stored at 4°C, we found that the CYP protein levels were unaffected by degradation after 72 h. At 21°C CYP1A2, CYP2D25, and CYP2E1 protein levels were nearly unaffected by degradation after 24 h, whereas a loss of approximately 50% was seen after 48 h. At 21°C CYP3A29 had a loss of 50% at 24 h and 70% at 48 h exhibiting less postmortem stability. In vitro enzyme activity measurements in the same tissue stored at 21°C showed a 50% decrease after 24 h and a complete loss of enzyme activity after 48 h. When stored at 4°C, the in vitro enzyme activity decreased to 50% activity after 96 h. In conclusion, measuring CYP levels by an LC–MS/MS approach was clearly less affected by postmortem changes than an activity‐based approach. The found postmortem stability for 24 h at 21°C for 3 out of 4 CYP isoforms supports the use of properly stored postmortem tissue to inform PBPK models.Kata W. PedersenJakob HansenJørgen B. HasselstrømJakob R. JornilWileyarticleCYP1A2CYP2D25CYP2E1CYP3A29cytochrome P450drug metabolismTherapeutics. PharmacologyRM1-950ENPharmacology Research & Perspectives, Vol 9, Iss 5, Pp n/a-n/a (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	CYP1A2 CYP2D25 CYP2E1 CYP3A29 cytochrome P450 drug metabolism Therapeutics. Pharmacology RM1-950
spellingShingle	CYP1A2 CYP2D25 CYP2E1 CYP3A29 cytochrome P450 drug metabolism Therapeutics. Pharmacology RM1-950 Kata W. Pedersen Jakob Hansen Jørgen B. Hasselstrøm Jakob R. Jornil Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification
description	Abstract Quantification of drug‐metabolizing cytochrome P450 (CYP) isoforms using LC–MS/MS has been proposed as a potential way of estimating antemortem CYP levels using postmortem tissue, but the postmortem stability of CYP proteins is incompletely investigated. If one can use data obtained from the analysis of postmortem specimens to inform physiologically based pharmacokinetic (PBPK) models this greatly increases the access to rare specimens among special subpopulations. In this study, we developed and validated an LC–MS/MS method for targeted CYP protein quantification in a porcine animal model to study postmortem stability. We measured 19.9–28.3 pmol CYP1A2, 50.3–66.2 pmol CYP2D25, 132.9–142.7 pmol CYP2E1, and 16.8–48 pmol CYP3A29 protein per mg PLM in nondegraded tissue. In tissue stored at 4°C, we found that the CYP protein levels were unaffected by degradation after 72 h. At 21°C CYP1A2, CYP2D25, and CYP2E1 protein levels were nearly unaffected by degradation after 24 h, whereas a loss of approximately 50% was seen after 48 h. At 21°C CYP3A29 had a loss of 50% at 24 h and 70% at 48 h exhibiting less postmortem stability. In vitro enzyme activity measurements in the same tissue stored at 21°C showed a 50% decrease after 24 h and a complete loss of enzyme activity after 48 h. When stored at 4°C, the in vitro enzyme activity decreased to 50% activity after 96 h. In conclusion, measuring CYP levels by an LC–MS/MS approach was clearly less affected by postmortem changes than an activity‐based approach. The found postmortem stability for 24 h at 21°C for 3 out of 4 CYP isoforms supports the use of properly stored postmortem tissue to inform PBPK models.
format	article
author	Kata W. Pedersen Jakob Hansen Jørgen B. Hasselstrøm Jakob R. Jornil
author_facet	Kata W. Pedersen Jakob Hansen Jørgen B. Hasselstrøm Jakob R. Jornil
author_sort	Kata W. Pedersen
title	Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification
title_short	Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification
title_full	Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification
title_fullStr	Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification
title_full_unstemmed	Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification
title_sort	stability investigations of cytochrome p450 (cyp) enzymes immediately after death in a pig model support the applicability of postmortem hepatic cyp quantification
publisher	Wiley
publishDate	2021
url	https://doaj.org/article/582ec299dd6a4b45822c779ccfcbe8c8
work_keys_str_mv	AT katawpedersen stabilityinvestigationsofcytochromep450cypenzymesimmediatelyafterdeathinapigmodelsupporttheapplicabilityofpostmortemhepaticcypquantification AT jakobhansen stabilityinvestigationsofcytochromep450cypenzymesimmediatelyafterdeathinapigmodelsupporttheapplicabilityofpostmortemhepaticcypquantification AT jørgenbhasselstrøm stabilityinvestigationsofcytochromep450cypenzymesimmediatelyafterdeathinapigmodelsupporttheapplicabilityofpostmortemhepaticcypquantification AT jakobrjornil stabilityinvestigationsofcytochromep450cypenzymesimmediatelyafterdeathinapigmodelsupporttheapplicabilityofpostmortemhepaticcypquantification
_version_	1718426481169793024

Stability investigations of cytochrome P450 (CYP) enzymes immediately after death in a pig model support the applicability of postmortem hepatic CYP quantification

Ejemplares similares