Lack of interaction between bropirimine and 5-fluorouracil on human dihydropyrimidine dehydrogenase

Abstract

1. Bropirimine (2-amino-5-bromo-6-phenyl-4-pyrimidinone) is a member of a class of antineoplastic agents that are administered concomitantly or sequentially with anticancer 5-fluorouracil (5-FU) prodrugs in clinical patients. Interactions between bropirimine and 5-fluorouracil (5-FU) were investigated on dihydropyrimidine dehydrogenase (DPD) activity, the rate-limiting enzyme of 5-FU metabolism, in human liver cytosol. Apparent DPD activity was determined by measuring the recovery of [¹⁴C]5-FU by HPLC.

2. The apparent activity of 5-FU metabolism (2.1 - 100 μm) showed a linear relationship in the Eadie-Hofstee plot in the pooled cytosol, suggesting that a single enzyme is responsible for apparent 5-FU metabolism. K_m and V_max were estimated to be 23 μm and 0.32 nmol min⁻¹ mg⁻¹ protein, respectively. Apparent DPD activity for 5-FU (25 μm) in the cytosol from 12 individual donors ranged from 0.017 to 0.39 (0.16 ± 0.12) nmol min⁻¹ mg⁻¹ protein, indicating a large intersubject variance.

3. The suicidal inactivators of the DPD enzyme, (E)-5-(2-bromovinyl)uracil and 5- bromouracil (6.3 - 50 μm), illustrated concentration-dependent inhibition on DPD activity. Isocytosine (6.3 - 100 μm), used as a negative control, did not affect DPD activity. Bropirimine (6.3 - 100 μm) also did not show any inhibition of DPD activity. Therefore, bropirimine is unlikely to cause increases in 5-FU levels in clinical patients after coadministration of bropirimine with 5-FU prodrugs.