Insights on cyclophosphamide metabolism and anticancer mechanism of action: A computational study

The oxazaphosphorine cyclophosphamide (CP) is a DNA-alkylating agent commonly used in cancer chemotherapy. This anticancer agent is administered as a prodrug activated by a liver cytochrome P450-catalyzed 4-hydroxylation reaction that yields the active, cytotoxic metabolite. The primary metabolite, 4-hydroxycyclophosphamide, equilibrates with the ring-open aldophosphamide that undergoes beta-elimination to yield the therapeutically active DNA cross-linking phosphoramide mustard and the byproduct acrolein. The present paper presents a DFT investigation of the different metabolic phases and an insight into the mechanism by which CP exerts its cytotoxic action. A detailed computational analysis of the energy profiles describing all the involved transformations and the mechanism of DNA alkylation is given with the aim to contribute to an increase of knowledge that, after more than 60 years of unsuccessful attempts, can lead to the design and development of a new generation of oxazaphosphorines. The outcomes of a density functional theory (DFT) computational mechanistic study of the main cyclophosphamide (CP) activation steps leading to the formation of the active phosphoramide mustard form and the DNA alkylating action of this latter are illustrated. Highlighting the kinetics and thermodynamics of each step can be helpful for the identification of appropriate structural modifications allowing CP improvement and/or the synthesis of more effective and less toxic analogues.image

Automated summary

This paper presents a DFT investigation on the metabolic phases and cytotoxic mechanism of cyclophosphamide (CP). The computational analysis provides insights into the energy profiles and DNA alkylation mechanism, aiming to contribute to the design of a new generation of oxazaphosphorines.