Anti‐fibrosis attributes: UHPLC‐MS/MS‐based pharmacokinetics profiling of a novel autotaxin inhibitor with excellent in vivo efficacy in rat.

3,4‐Difluorobenzyl(1‐ethyl‐5‐(4‐((4‐hydroxypiperidin‐1‐yl)‐methyl)thiazol‐2‐yl)‐1H‐indol‐3‐yl)carbamate (NAI59), a small molecule with outstanding therapeutic effectiveness to anti‐pulmonary fibrosis, was developed as an autotaxin inhibitor candidate compound. To evaluate the pharmacokinetics and plasma protein binding of NAI59, a UPLC–MS/MS method was developed to quantify NAI59 in plasma and phosphate‐buffered saline. The calibration curve linearity ranged from 9.95 to 1990.00 ng/mL in plasma. The accuracy was −6.8 to 5.9%, and the intra‐ and inter‐day precision was within 15%. The matrix effect and recovery, as well as dilution integrity, were within the criteria. The chromatographic and mass spectrometric conditions were also feasible to determine phosphate‐buffered saline samples, and it has been proved that this method exhibits good precision and accuracy in the range of 9.95–497.50 ng/mL in phosphate‐buffered saline. This study is the first to determine the pharmacokinetics, absolute bioavailability, and plasma protein binding of NAI59 in rats using this established method. Therefore, the pharmacokinetic profiles of NAI59 showed a dose‐dependent relationship after oral administration, and the absolute bioavailability in rats was 6.3%. In addition, the results of protein binding showed that the combining capacity of NAI59 with plasma protein attained 90% and increased with the increase in drug concentration. [ABSTRACT FROM AUTHOR]