Determination of A Novel Antifibrotic Small Molecule GDC-3280 in Human Plasma and Urine by Liquid Chromatography Tandem Mass Spectrometry to Support Its First-in-human Clinical Trial.

A specific and robust LC-MS/MS method was developed and validated for the quantitative determination of GDC-3280 in human plasma and urine. The nonspecific binding associated with urine samples was overcome by the addition of CHAPS. The sample volume was 25 μL for either matrix, and supported liquid extraction was employed for analyte extraction. d6-GDC-3280 was used as the internal standard. Linear standard curves (R2 >0.9956) were established from 5.00 to 5000 ng/mL in both matrices with quantitation extended to 50,000 ng/mL through dilution. In plasma matrix, the precision (%RSD) ranged from 1.5% to 9.9% (intra-run) and from 2.4% to 7.2% (inter-run); the accuracy (%RE) ranged from 96.1% to 107% (intra-run) and from 96.7% to 104% (inter-run). Similarly, in urine the precision was 1.5% to 6.2% (intra-run) and 1.9% to 6.1% (inter-run); the accuracy was 83,1% to 99.3% (intra-run) and 87.1% to 98.3% (inter-run). Good recovery (>94%) and negligible matrix effect were achieved in both matrices. Long-term matrix stability was established for at least 703 days in plasma and 477 days in urine. Benchtop stability of 25 hrs and 5 freeze-thaw cycles were also confirmed in both matrices. The method was successfully implemented in GDC-3280's FIH trial for assessing its pharmacokinetic profiles.