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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

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Abstract

LJ-2698, a highly potent human A3 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.

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Acknowledgements

This work was supported by the Basic Study for Well Aging Project funded by the Seokchun Daewoong Foundation (2015).

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Authors and Affiliations

  1. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, 08826, Republic of Korea

    Jae-Young Lee, Ju-Hwan Park, Ki-Taek Kim, Jinha Yu, Pramod K. Sahu, Naewon Kang, Hyeon-Jong Shin, Min-Hwan Kim, Ji-Su Kim, Lak Shin Jeong & Dae-Duk Kim

  2. College of Pharmacy, Chungnam National University, Daejeon, 34134, Republic of Korea

    Jae-Young Lee

  3. Future Medicine Co., Ltd, Seoul, 06665, Republic of Korea

    Pramod K. Sahu

  4. College of Pharmacy and Natural Medicine Research Institute, Mokpo National University, Jeonnam, 58628, Republic of Korea

    In-Soo Yoon

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  1. Jae-Young Lee
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Lee, JY., Park, JH., Kim, KT. et al. 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. Arch. Pharm. Res. 40, 952–961 (2017). https://doi.org/10.1007/s12272-017-0935-9

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