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 (10 mg/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 ± 5.8 L/h/kg), high volume of distribution (244 ± 51.5 L/kg) and a terminal half-life of 4.8 ± 1.6 h. 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.

Development of a validated LC–MS/MS method for the determination of ailanthone in rat plasma with application to pharmacokinetic study

“…The pharmacokinetic parameters of the three dosages calculated using noncompartmental analysis were shown in Table 3. In this study, it can be seen that the concentration of ailanthone in rat plasma after intravenous administration was relatively high although the dosages were lower than some other PK researches [19][20][21], which might be due to the good solubility of ailanthone. The t 1/2 were 105.5 ± 13.6, 113.3 ± 39.6 and 95.8 ± 23.9 min after administration of 0.5, 1.0, and 1.5 mg/kg ailanthone, respectively, which means that ailanthone is eliminated fast in rats.…”

Discoveries of Targets and Novel Agents for the Treatment of Ischemic Retinopathy and Neovascular Disease

“…Murine ocular pharmacokinetics of KZ-41 delivered topically to the mouse eye was characterized for penetration into ocular tissues using analytical methods described previously [166,167]. Adult C57BL/6J mice received a single ocular administration of KZ-41-loaded NE (100 mg/kg) to the right eye; the left eye was dosed with vehicle for contralateral controls.…”

“…Pharmacokinetic studies were performed as previously described [166,167,253,254]. Animals will receive either a single intravenous or oral dose of JP-153 (5, 10 mg/kg, respectively).…”