Synthesis of cyclophosphamide metabolites by a peroxygenase from Marasmius rotula for toxicological studies on human cancer cells.
Biocatalysis
Cyclophosphamide
Human drug metabolites
Peroxygenase
Toxicity
Journal
AMB Express
ISSN: 2191-0855
Titre abrégé: AMB Express
Pays: Germany
ID NLM: 101561785
Informations de publication
Date de publication:
18 Jul 2020
18 Jul 2020
Historique:
received:
26
06
2020
accepted:
13
07
2020
entrez:
20
7
2020
pubmed:
20
7
2020
medline:
20
7
2020
Statut:
epublish
Résumé
Cyclophosphamide (CPA) represents a widely used anti-cancer prodrug that is converted by liver cytochrome P450 (CYP) enzymes into the primary metabolite 4-hydroxycyclophosphamide (4-OH-CPA), followed by non-enzymatic generation of the bioactive metabolites phosphoramide mustard and acrolein. The use of human drug metabolites as authentic standards to evaluate their toxicity is essential for drug development. However, the chemical synthesis of 4-OH-CPA is complex and leads to only low yields and undesired side products. In past years, fungal unspecific peroxygenases (UPOs) have raised to powerful biocatalysts. They can exert the identical selective oxyfunctionalization of organic compounds and drugs as known for CYP enzymes with hydrogen peroxide being used as sole cosubstrate. Herein, we report the efficient enzymatic hydroxylation of CPA using the unspecific peroxygenase from Marasmius rotula (MroUPO) in a simple reaction design. Depending on the conditions used the primary liver metabolite 4-OH-CPA, its tautomer aldophosphamide (APA) and the overoxidized product 4-ketocyclophosphamide (4-keto-CPA) could be obtained. Using a kinetically controlled approach 4-OH-CPA was isolated with a yield of 32% (purity > 97.6%). Two human cancer cell lines (HepG2 and MCF-7) were treated with purified 4-OH-CPA produced by MroUPO (4-OH-CPA
Identifiants
pubmed: 32683510
doi: 10.1186/s13568-020-01064-w
pii: 10.1186/s13568-020-01064-w
pmc: PMC7368878
doi:
Types de publication
Journal Article
Langues
eng
Pagination
128Subventions
Organisme : Ministerium für Wissenschaft, Forschung und Kultur
ID : 22-F241-03-FhG/005/001
Organisme : European Regional Development Fund
ID : 85002925
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