title: Effect of nitroreduction on the alkylating reactivity and cytotoxicity of the 2,4-dinitrobenzamide-5-aziridine CB 1954 and the corresponding nitrogen mustard SN 23862: distinct mechanisms of bioreductive activation.
abstract: The dinitrobenzamide aziridine CB 1954 (1) and its nitrogen mustard analogue SN 23862 (6) are prodrugs that are activated by enzymatic nitroreduction in tumors. Bioactivation of 1 is considered to be due to reduction of its 4-nitro group to the hydroxylamine and subsequent formation of the N-acetoxy derivative; this acts as a reactive center, in concert with the aziridine moiety, to provide a bifunctional DNA cross-linking agent (Knox model). It is currently unclear whether bioactivation of 6 occurs by the same mechanism or results from the electronic effects of nitroreduction on reactivity of the nitrogen mustard moiety. To discriminate between these mechanisms, we have synthesized the hydroxylamine and amine derivatives of 1 and 6, plus related compounds, and determined their alkylating reactivities in aqueous solution, using LC/MS to identify reaction pathways. The relationships between substituent electronic effects, reactivity, and cytotoxicity were determined using the UV4 cell line, which is defective in nucleotide excision repair (thus avoiding differences in repair kinetics). Alkylating reactivity correlated with the electron-donating character of the ortho or para substituent in the case of the mustards, with a less marked electronic effect for the aziridines. Importantly, there was a highly significant linear relationship between cytotoxic potency and alkylating reactivity in both the aziridine and the mustard series, with the notable exception of 4, the 4-hydroxylamine of 1, which was 300-fold more toxic than predicted by this relationship. This demonstrates that the high potency of 4 does not result from activation of the aziridine ring, supporting the Knox model. The single-step bioactivation of 6, to amino or hydroxylamine metabolites with similar potency to 4, is a potential advantage in the use of dinitrobenzamide mustards as prodrugs for activation by nitroreductases.
authors.affiliation: Auckland Cancer Society Research Centre, The University of Auckland, Private Bag 92019, Auckland, New Zealand. n.helsby@auckland.ac.nz
authors.forename: Nuala A
authors.initials: NA
authors.lastname: Helsby
authors.forename: S James
authors.initials: SJ
authors.lastname: Wheeler
authors.forename: Frederik B
authors.initials: FB
authors.lastname: Pruijn
authors.forename: Brian D
authors.initials: BD
authors.lastname: Palmer
authors.forename: Shangjin
authors.initials: S
authors.lastname: Yang
authors.forename: William A
authors.initials: WA
authors.lastname: Denny
authors.forename: William R
authors.initials: WR
authors.lastname: Wilson
mesh_terms: D000816:Aniline Mustard / Q000031:analogs & derivatives* / Q000378:metabolism*; D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000378:metabolism*; D001388:Aziridines / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004563:Electrochemistry; D006868:Hydrolysis; D007700:Kinetics; D013058:Mass Spectrometry; D009601:Nitroreductases / Q000378:metabolism*; D011355:Prodrugs / Q000378:metabolism*; D013997:Time Factors; D014407:Tumor Cells, Cultured
chemical_list: D018906:Antineoplastic Agents, Alkylating; D001388:Aziridines; D011355:Prodrugs; C076205:5-(N,N-bis(2-chloroethyl)amino)-2,4-dinitrobenzamide; C100099:tretazicar; D000816:Aniline Mustard; D009601:Nitroreductases