Mutagenic, teratogenic and pharmacokinetic properties of cyclophosphamide and some of its deuterated derivatives

Abstract

To elucidate which metabolic pathway leads to the ultimate mutagenic and teratogenic metabolite of cyclophosphamide (CPA), the mutagenicity in vitro as well as the teratogenicity in vivo and the pharmacokinetics of CPA and several deuterated analogs (5,5-d 2-CPA, 4,4-d 2-CPA and 4,4–6,6-d 4-CPA) were compared. 5,5-d 2-CPA was less mutagenic (isotope effects between 1.7 and 12.3 were found for sister-chromatid exchanges and structural chromosomal aberrations in CHO cells and in the Ames test) and less teratogenic (deuterium isotope effect between and 3) than CPA and the other deuterated analogs. Because the concentrations of 5,5-d 2-CPA in pregnant mice and their embryos were equal to or even exceeded those of CPA and the other deuterated analogs, a particular metabolic pathway involving a fission of the C-D or C-H bond on the C-5 rather than the parent drug must be responsible for the isotope effect observed. It had previously been shown (Cox et al., 1976) that the 5,5-d 2-CPA has a decreased anti-tumor activity in vivo (isotope effect 7–13) which was accompanied by a decreased formation of phosphoramide mustard and acrolein in vitro (isotope effect 5.3). Our results suggest that phosphoramide mustard is an important proximate metabolite of CPA with regard to the mutagenicity and teratogenicity of the drug. These results show for the first time that labeling of drugs with stable isotopes offers a promising approach to the study of the mechanism of the mutagenic and teratogenic actions of drugs.