Determination and validation of LJ-2698, a potent human A3 adenosine receptor antagonist, in rat plasma by liquid chromatography-tandem mass spectrometry and its application in pharmacokinetic study

• Published in: Archives of pharmacal research Vol. 40; no. 8; pp. 952 - 961
• Main Authors: Lee, Jae-Young, Park, Ju-Hwan, Kim, Ki-Taek, Yu, Jinha, Sahu, Pramod K., Kang, Naewon, Shin, Hyeon-Jong, Kim, Min-Hwan, Kim, Ji-Su, Yoon, In-Soo, Jeong, Lak Shin, Kim, Dae-Duk
• Format: Journal Article
• Language: English
• Published: Seoul Pharmaceutical Society of Korea 01.08.2017; 대한약학회
• Subjects: acetonitrile; antagonists; dose response; formic acid; humans; intravenous injection; ionization; liquid chromatography; Medicine; monitoring; nucleosides; oral administration; pharmacokinetics; Pharmacology/Toxicology; Pharmacy; purinergic receptors; rats; Research Article; statistical analysis; tandem mass spectrometry; 약학; Validation; LJ-2698; Adenosine analogues; AR antagonist; hA; Pharmacokinetics; LC–MS/MS
• ISSN: 0253-6269; 1976-3786; 1976-3786
• DOI: 10.1007/s12272-017-0935-9

LJ-2698, a highly potent human A 3 adenosine receptor antagonist with nucleoside structure, was designed to have a minimal species dependence. For further pre-clinical studies, analytical method for the detection of LJ-2698 in rat plasma was developed by liquid chromatography-tandem mass. Plasma samples were processed by protein precipitation method with acetonitrile, using losartan as the internal standard (IS). Chromatographic separation was carried out using a Kinetex C18 column (100 × 4.6 mm; 100 Å; 2.6 μ) with acetonitrile/water with 0.2% (v/v) formic acid (65:35, v/v) in the isocratic mode at a flow rate of 0.4 mL/min. Mass spectrometric detection in multiple reaction monitoring mode was performed with positive electrospray ionization. The mass transitions of LJ-2698 and IS were m/z 412.3 → 294.1 and m/z 423.1 → 207.2, respectively. The calibration curves were linear in the range 5.00–5000 ng/mL ( r 2  ≥ 0.998). The lower limit of quantification was established as 5.00 ng/mL. Within- and between-run precisions were <7.01%, as relative standard deviation; and accuracies were in the range 3.37–3.64%, as relative error. The validated method was successfully applied to its pharmacokinetic evaluation after intravenous and oral administration in rats, and the dose-dependent pharmacokinetic behavior of LJ-2698 was elucidated for the first time.