Pharmacokinetic and pharmacodynamic assessments
PD
BLOOD
Blood samples (4 mL) for PK analysis were obtained in chilled collection tubes containing the anticoagulant KThe plasma samples were analysed for TQA3526 using a validated analytical method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) using human plasma, urine, and faecal samples. The linearity ranges were 0.2–200 ng/mL for TQA3526 in the plasma. The precision rate was ≤5.7%, with an accuracy in the range of −0.6 to 1.7%.ALP, FGF19, bile acid precursor C4 (7α-hydroxy-cholest-4-ene-3-one), cholesterol, and bile acid were selected for PD analysis. PD blood samples were obtained at 0 h (pre-dose), and 2 h, 4 h, 6 h, 8 h, 12 h (D1), 24 h (D2), 72 h (D4) in parts 1 and 3. For part 2, the ninth cohort, blood samples were obtained at 0 h (pre-dose), and 2 h, 4 h, 6 h, 8 h, 12 h (D1 and D7), D2, D3, D5, D8, and D9. In the tenth cohort, FGF19 was obtained at 0 h (pre-dose), and 10 min, 20 min, 30 min, 45 min, 1 h, 1.5 h, 2 h, 3 h, 4 h, 6 h, 8 h, 12 h (D20 and D30), D8–D10, D15, D17, D19, D21, D25, D27, D29, and D31–D33.Serum ALP levels were measured using a Beckman Coulter kit assay with neryl diphosphate (NPP) as substrates and 2-amino-2-methyl-1-propanol (AMP) as buffers. Cholesterol activity was measured using a Beckman Coulter test kit with enzyme method. Total bile acid activity was measured using a Beckman Coulter kit with circulating enzymatic method. FGF-19 was tested using a quantification assay by an ELISA kit. C4 was analysed using a validated analytical method based on LC-MS/MS.