Development, Validation of LC-MS/MS Method and Determination of Pharmacokinetic Parameters of the Stroke Neuroprotectant Neurounina-1 in Beagle Dog Plasma After Intravenous Administration

Author(s):Show less -

Severino, Beatrice ^[1] ; Corvino, Angela ^[1] ; Fiorino, Ferdinando ^[1] ; Frecentese, Francesco ^[1] ; Perissutti, Elisa ^[1] ; Caliendo, Giuseppe ^[1] ; Santagada, Vincenzo ^[1] ; Magli, Elisa ^[1] ; Molinaro, Pasquale ^[2] ; Pignataro, Giuseppe ^[2] ; Annunziato, Lucio ^[3] ; Antunes, Natalicia J. ^[4] ; Rojas-Moscoso, Julio ^[4] ; de Freitas, Noedi L. ^[4] ; Mendes, Gustavo D. ^{[5, 6, 7]} ; De Nucci, Gilberto ^{[4, 8, 5]}

Affiliation

^[1] Univ Naples Federico II, Sch Med, Dept Pharm, Naples - Italy

^[2] Univ Naples Federico II, Sch Med, Dept Neurosci Reprod & Odontostomatol Sci, Naples - Italy

^[3] IRCCS SDN, Naples - Italy

^[4] Univ Estadual Campinas, Fac Med Sci, Dept Pharmacol, Campinas, SP - Brazil

^[5] Sao Leopoldo Mandic, Fac Med, Dept Pharmacol, Campinas, SP - Brazil

^[6] Univ Metropolitana Santos, Fac Med Sci, Dept Pharmacol, Santos - Brazil

^[7] Univ Mogi das Cruzes, Fac Med, Mogi Das Cruzes - Brazil

^[8] Univ Sao Paulo, Fac Med Sci, Dept Pharmacol, Sao Paulo - Brazil

• Document type: Journal article
• Source: FRONTIERS IN PHARMACOLOGY; v. 10, APR 25 2019.
• Web of Science Citations: 1

Abstract

Neurounina-1 {[}chemical name: 7-nitro-5-phenyl-1-(pyrrolidin-1-ylmethyl)-1H-benzo{[}e]{[}1,4]diazepin- 2(3H)-one] is a new compound provided with relevant neuroprotective effect during stroke and in neonatal hypoxia by increasing the Na+/Ca2+ exchanger (NCX) isoforms NCX1 and NCX2 activity. This study shows for the first time, the development and validation of a sensitive and selective method for analysis of neurounina-1 in beagle dog plasma by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The sample preparation consisted of extraction of the analyte and the internal standard (IS) (ropivacaine) from plasma (50 mu L) by liquid-liquid extraction using acetonitrile (100 mu L). The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 365 > 83 and m/z 275 > 126 were used to measure the derivative of neurounina-1 and ropivacaine. The chromatographic separation was achieved using a Phenomenex C18 Luna (150 mm x 4.6 mm x 5 mu m) analytical column with an isocratic mobile phase composed of methanol/acetonitrile/water (50/40/10, v/v/v) + 0.1% formic acid + 1 M ammonium formate. The method was linear over a concentration range of 1-500 ng/mL. The method was applied to evaluate the pharmacokinetics of neurounina-1 after a single intravenous administration of three different doses (0.1 mg/kg, 0.3 mg/kg, and 1 mg/kg) to beagle dogs (n = 5). The mean AUC(0-tlast) values were 26.10, 115.81, and 257.28 ng{*}h/mL following intravenous administration of 0.1, 0.3, and 1 mg/kg, respectively. Linear pharmacokinetics was observed up to 1.0 mg/kg. The neurounina-1 was rapidly eliminated, with mean CL values of 46.24, 47.57, and 69.15 L/h, Vd of 130.31, 154.15, and 210.79 L and t(1/2) of 2.14, 2.54, and 2.04 h after intravenous administration of 0.1, 0.3, and 1 mg/kg, respectively. This new analytical method allows the rapid determination of the neurounina-1, a new developed compound, able to exert a remarkable neuroprotective effect in the low nanomolar range. (AU)

• FAPESP's process: 16/22506-1 - Metabolism of dapaconazole
• Grantee: Natalícia de Jesus Antunes
• Support Opportunities: Scholarships in Brazil - Post-Doctoral