Figure 1.
Identification of belinostat metabolites in human plasma using HPLC-UV at maximum absorption wavelength (λ = 268 nm).
Chromatogram at day 5 and day 22 (A); day1(B).
Table 1.
Pharmacokinetic parameters for belinostat after 30 min i.v. infusion in Phase I trial.
Figure 2.
Belinostat metabolism pathway in human plasma with glucuronidation of belinostat as the dominant metabolism.
Table 2.
Identification of belinostat metabolites in human plasma using HPLC-DAD & LC-MS/MS analyses.
Figure 3.
Cytotoxicity and acetylation activity on HepG2.
a: belinostat incubation; b: belinostat-G incubation; (templates for concentrations added (lower) and results of 24-well dose-increasing concentrations on HepG2 Cells (upper). c: MTS results for belinostat (IC50 = 6.4 µM) and belinostat-G (cannot be converged). d:Belinostat acetylation activity on HepG2 cells (western blot). A: Acetyl histone 3 increased with dose increment after 5 h incubation; B: Kinetic changes of acetyl histone 3 with time increment at 10 µM.
Figure 4.
Enzyme stability test in a panel of 14 UGT isoforms after 2 h incubation at 37°C (A); Time course of glucuronidation of belinostat by UGT1A1supersomes at 37°C (B).
Figure 5.
Enzyme kinetics of glucuronidation of belinostat by UGT1A1.
The apparent Km and Vmax values for the glucuronide formation were 99.6 µM and 353.1 pmol/min/mg protein, respectively.
Figure 6.
Association between glucuronidation of belinostat and UGT1A1 substrates (A, B, and C).
A: Bilirubin-G; B: thyroxine-4-G/CPT-11; C: SN38-G/CPT11.
Figure 7.
UGT1A1 expression on belinostat glucuronidation and impact of the common UGT1A1*28 promoter polymorphism.
A: Correlation of belinostat glucuronide formation with UGT1A1 expression in human liver microsomes; B: Belinostat glucuronide formation by human liver microsomes according to wild-type, heterozygous and homozygous UGT1A1*28 genotypes.