title: A sensitive and fast LC-MS/MS method for determination of β-receptor agonist JP-49b: application to a pharmacokinetic study in rats.
abstract: Ocular administration of the beta (β)-adrenergic receptor agonist JP-49b prevents retinopathy-like damage in a preclinical rat model of diabetes. Importantly, JP-49b did not induce characteristic β-adrenergic agonist-related side effects (e.g., left ventricular damage), which led to the hypothesis that JP-49b systemic exposure was minimal following ocular administration. To test this hypothesis, a sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to study the preclinical pharmacokinetics of JP-49b in rats. Animals received either a single periocular or intravenous injection of JP-49b (10mg/kg) and plasma and tissue samples were obtained. JP-49b and fenoterol hydrobromide (internal standard, IS) were isolated by liquid-liquid extraction and extracts were analyzed by reversed-phase liquid chromatography on a C18 column using a gradient elution (acetic acid in water and methanol). A triple quadrupole mass spectrometer operating in the positive electrospray ionization mode with multiple reaction monitoring was used to detect JP-49b and IS transitions of m/z 346.4→195.1 and 304.1→134.9. The method was validated for selectivity, linearity, accuracy, and precision in rat vitreous humor, tissue homogenates, and plasma. Following intravenous administration, JP-49b was found to have a rapid clearance (36±5.8L/h/kg), high volume of distribution (244±51.5L/kg) and a terminal half-life of 4.8±1.6h. JP-49b was rapidly absorbed and extensively distributed into ocular tissue following topical administration. However, JP-49b was undetectable in heart tissue 24h after ocular administration. High local drug concentrations coupled with minimal systemic exposure following ocular administration supports further testing of JP-49b as a localized therapy for diabetic retinopathy.
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, United States.
authors.forename: Hui
authors.initials: H
authors.lastname: He
authors.affiliation: Department of Ophthalmology, University of Tennessee Health Science Center, Memphis, TN 38163, United States.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Williams-Guy
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, United States.
authors.forename: Jayaprakash
authors.initials: J
authors.lastname: Pagadala
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, United States.
authors.forename: Chaela Sickbert
authors.initials: CS
authors.lastname: Presley
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, United States.
authors.forename: Duane D
authors.initials: DD
authors.lastname: Miller
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, United States; Department of Ophthalmology, University of Tennessee Health Science Center, Memphis, TN 38163, United States; Department of Anatomy and Neurobiology, University of Tennessee Health Science Center, Memphis, TN 38163, United States.
authors.forename: Jena J
authors.initials: JJ
authors.lastname: Steinle
authors.affiliation: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, United States; Department of Ophthalmology, University of Tennessee Health Science Center, Memphis, TN 38163, United States. Electronic address: cyates4@uthsc.edu.
authors.forename: Charles R
authors.initials: CR
authors.lastname: Yates
mesh_terms: D000318:Adrenergic beta-Agonists / Q000008:administration & dosage / Q000032:analysis / Q000097:blood* / Q000493:pharmacokinetics*; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D056965:Injections, Intraocular; D007275:Injections, Intravenous; D016014:Linear Models; D008297:Male; D009638:Norepinephrine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000493:pharmacokinetics; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014822:Vitreous Body / Q000737:chemistry
chemical_list: D000318:Adrenergic beta-Agonists; C000592881:JP-49b; D009638:Norepinephrine
keywords: JP-49b; LC–MS/MS; Rat pharmacokinetics; β-receptor agonist