Interspecies comparison of plasma metabolism and sample stabilization for quantitative bioanalyses: Application to (R)-CE3F4 in preclinical development, including metabolite identification by high-resolution mass spectrometry.

Autor: Toussaint B; Université Paris-Saclay, CNRS, Institut Galien Paris Sud, 92296 Châtenay-Malabry, France; Département de Recherche et Développement Pharmaceutique, Agence Générale des Équipements et Produits de Santé (AGEPS), Assistance Publique des Hôpitaux de Paris (AP-HP), Paris, France., Hillaireau H; Université Paris-Saclay, CNRS, Institut Galien Paris Sud, 92296 Châtenay-Malabry, France., Jaccoulet E; Université Paris-Saclay, CNRS, Institut Galien Paris Sud, 92296 Châtenay-Malabry, France; Hôpital européen Georges Pompidou (HEGP), Service Pharmacie (AP-HP), Paris, France., Cailleau C; Université Paris-Saclay, CNRS, Institut Galien Paris Sud, 92296 Châtenay-Malabry, France., Legrand P; Département de Recherche et Développement Pharmaceutique, Agence Générale des Équipements et Produits de Santé (AGEPS), Assistance Publique des Hôpitaux de Paris (AP-HP), Paris, France; Université de Paris, Faculté de sciences pharmaceutiques et biologiques, Unité de Technologies Chimiques et Biologiques pour la Santé (UTCBS), CNRS UMR8258, Inserm U1022, Paris, France., Ambroise Y; Université Paris-Saclay, CEA, Institut des Sciences du Vivant Frederic Joliot, 91191 Gif-sur-Yvette, France., Fattal E; Université Paris-Saclay, CNRS, Institut Galien Paris Sud, 92296 Châtenay-Malabry, France. Electronic address: elias.fattal@universite-paris-saclay.fr.
Jazyk: angličtina
Zdroj: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2021 Oct 15; Vol. 1183, pp. 122943. Date of Electronic Publication: 2021 Sep 30.
DOI: 10.1016/j.jchromb.2021.122943
Abstrakt: The CE3F4 is an inhibitor of the type 1 exchange protein directly activated by cAMP (EPAC1), which is involved in numerous signaling pathways. The inhibition of EPAC1 shows promising results in vitro and in vivo in different cardiac pathological situations like hypertrophic signaling, contributing to heart failure, or arrhythmia. An HPLC-UV method with a simple and fast sample treatment allowed the quantification of (R)-CE3F4. Sample treatment consisted of simple protein precipitation with 50 µL of ethanol and 150 µL of acetonitrile for a 50 µL biological sample. Two wavelengths were used according to the origin of plasma (220 or 250 nm for human samples and 250 nm for murine samples). Accuracy profile was evaluated for both wavelengths, and the method was in agreement with the criteria given by the EMA in the guideline for bioanalytical method validation for human and mouse plasma samples. The run time was 12 min allowing the detection of the (R)-CE3F4 and a metabolite. This study further permitted understanding the behavior of CE3F4 in plasma by highlighting an important difference between humans and rodents on plasma metabolism and may impact future in vivo studies related to this molecule and translation of results between animal models and humans. Using paraoxon as a metabolism inhibitor was crucial for the stabilization of (R)-CE3F4 in murine samples. HPLC-UV and HPLC-MS/MS studies were conducted to confirm metabolite structure and consequently, the main metabolic pathway in murine plasma.
(Copyright © 2021. Published by Elsevier B.V.)
Databáze: MEDLINE
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