Accelerated decomposition of 4-hydroxycyclophosphamide by human serum albumin.
Cyclophosphamide, a widely used anticancer agent, requires initial metabolic activation to 4-hydroxycyclophosphamide (4-OHCP) to elicit its activity. The rate of decomposition of cis-4-OHCP was much faster in plasma than in buffer at pH 7.4. This plasma activity was not affected by treatment with acid (pH 1.3) or heat (60 degrees C for 30 min). The activity was retained in the macromolecular fraction (greater than 10,000) but not in the filtrate. Serum albumin was identified as the catalyst for the elimination step that generates phosphoramide mustard from aldophosphamide; albumin had no effect on the rate of ring opening of cis-4-OHCP to aldophosphamide. This catalytic activity was dependent on serum albumin concentration and independent of pH over the range of 6.5 to 7.5, in contrast to the buffer-catalyzed reaction. The catalytic rate constants kcat (pH 7.4, 37 degrees C) for phosphate buffer, human serum albumin, and bovine serum albumin were 1.13, 285, and 83 M-1 min-1, respectively. Pretreatment of cis-4-OHCP with serum albumin resulted in a time-dependent decrease in cytotoxic activity against L1210 tumor cells in vitro. These data suggest that the albumin-catalyzed reaction of cis-4-OHCP in plasma represents an important pathway for the transformation of cyclophosphamide metabolites and further emphasize the importance of considering phosphoramide mustard generated extracellularly versus intracellularly and the respective contributions of extracellular and intracellular phosphoramide mustard to cyclophosphamide cytotoxicity in vivo.
D002470:Cell Survival / Q000187:drug effects; D003520:Cyclophosphamide / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D010752:Phosphoramide Mustards / Q000378:metabolism; D012709:Serum Albumin / Q000494:pharmacology*
D010752:Phosphoramide Mustards; D012709:Serum Albumin; C030090:phosphoramide mustard; C012358:4-hydroxycyclophosphamide; D003520:Cyclophosphamide