The Open Drug Metabolism Journal
2010, 4 : 1-9Published online 2010 January 27. DOI: 10.2174/1874073101004010001
Publisher ID: TODMJ-4-1
RESEARCH ARTICLE
In Vitro Profiling and Mass Balance of the Anti-Cancer Agent Laromustine [C]-VNP40101M by Rat, Dog, Monkey and Human Liver Microsomes
* Address correspondence to this author at the Vion Pharmaceuticals, Inc., Four Science Park, New Haven, CT 06511, USA; Tel: 203-672-4561; Fax: 203-781-8090; E-mail: nassaral@aol.com
ABSTRACT
Laromustine (VNP40101M; 1,2-bis(methylsulfonyl)-1-(2-chloroethyl)-2-(methylamino) carbonylhydrazine) is a novel sulfonylhydrazine alkylating agent. For the first time In vitro profiling and mass balance of [14C] VNP40101M in rat, dog, monkey and human liver microsomes was investigated. Also, the role of human cytochrome P450 (CYP) and flavin-containing monooxygenase (FMO) enzymes in the conversion of [14C] VNP40101M by NADPH-fortified human liver microsomes was determined. In this study, [14C]-VNP40101M was converted to five radioactive components (C-1, C-2, C-3, C-4 and C-7) after 60 min of incubation with dog, monkey and human liver microsomes. With the exception of C-3, the same components were detected with rat liver microsomes. In the presence of NADPH, after 60 min of incubation, the loss of substrate for rat, dog, monkey and human was 63, 82, 76 and 64%, respectively and mass balance ranged from 91.0 – 99.3%. In the absence of NADPH, after 60 min of incubation with [14C] VNP40101M (100 µM), the loss of substrate for rat, dog, monkey and human liver microsomes was 59, 53, 61 and 59%, respectively and mass balance ranged from 100.6 – 116.4%. The profiles of metabolites were similar. The relative abundance of individual metabolites was not species dependent. The formation of C-7 was not observed in zero cofactor (no NADPH) or zero-protein samples, suggesting that its formation was enzymatic. The formation of C-1, C-2, C-3, and C-4 increased with respect to incubation time. Using a panel of CYP enzymes including rCYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6 and 3A4, it was shown that C-7 formation was catalyzed by CYP2B6 and CYP3A4/5. The results of this study suggest that (1) P450 plays a role in C-7 formation but plays little or no role in the conversion of [14C] VNP40101M to C-1 through C-4, and (2) the relative abundance of individual degradation/metabolite products were not species dependent. These findings provide a comprehensive understanding of the metabolism of this new agent.