Hypoxia-selective antitumor agents. 3. Relationships between structure and cytotoxicity against cultured tumor cells for substituted N,N-bis(2-chloroethyl)anilines

Abstract

A series of aniline mustards with a wide range of electron-donating and -withdrawing substituents in the 3- and 4-positions has been synthesized and evaluated for cytotoxicity in cell culture to examine the potential of using nitro group deactivated nitrogen mustards for the design of novel hypoxia-selective anticancer drugs (Denny, W. A.; Wilson, W. R. J. Med. Chem. 1986, 29, 879). Hydrolytic half-lives in tissue culture media, determined by bioassay against a cell line (UV4) defective in the repair of DNA interstrand cross-links showed the expected dependence on the Hammett electronic parameter, sigma, varying from 0.13 h for the 4-amino analogue to greater than 100 h for analogues with strongly electron-withdrawing substituents. Cytotoxic potencies in aerobic UV4 cultures showed a similar dependence on sigma. This dependence predicted that the 4-nitroaniline mustard would be 7200-fold less potent than its potential six-electron reduction product, the 4-amino compound, in growth inhibition assays using a 1-h drug exposure. The measured differential was much lower (225-fold) because of the instability of the latter compound, but a differential of 17,500-fold was observed in the initial rate of killing by using a clonogenic assay. The potential for formation of reactive mustards by reduction to the amine or hydroxylamine was demonstrated by the 4-nitroso compound, which had an aerobic toxicity similar to that of the amine. Although these features confirmed the original rationale, the 3-nitro- and 4-nitroaniline mustards had only minimal hypoxic selectivity against UV cells. Toxicity to hypoxic cells appears to be limited by the low reduction potentials of these compounds and consequent lack of enzymatic nitroreduction. However, this study has demonstrated that nitro groups can be used to latentiate aromatic nitrogen mustards and indicates that examples with higher reduction potentials could provide useful hypoxia-selective therapeutic agents.

Hypoxia-Activated Prodrugs in Cancer Therapy: Progress to the Clinic

“…The reactivity and cytotoxicity of aromatic nitrogen mustards is known to depend largely on the electron den� sity at the mustard nitrogen, with very large differences in cytotoxicity observed between nitroaromatic mustards and their four�electron (hydroxylamine) or 6�electron (amine) products (Figure 2A) [43] .…”

Influence of Mustard Group Structure on Pathways of in Vitro Metabolism of Anticancer N-(2-Hydroxyethyl)-3,5-dinitrobenzamide 2-Mustard Prodrugs

“…Enzymatic nitroreduction converts an electron-withdrawing nitro group ( p ϩ0.78) to a hydroxylamine ( p Ϫ0.32) or amine ( p Ϫ0.66) metabolite; this "electronic switch" can be used to activate the latent nitrogen mustard moiety (Denny and Wilson, 1986;Palmer et al, 1990;Siim et al, 1997) The 2,4-dinitrobenzamide-5-mustard class (2,4-DNBM) was investigated first and shown to provide cytotoxicity and DNA cross-linking selectively under hypoxic conditions (Palmer et al, 1992). The 2,4-DNBMs were subsequently shown to be substrates for the oxygen-insensitive nitroreductase nfsB from Escherichia coli (Anlezark et al, 1995;Wilson et al, 2002;Atwell et al, 2007), which is of interest for prodrug activation in the context of gene-or virus-directed enzyme prodrug therapy.…”

Potentiation of antitumor immunity by antibody-directed enzyme prodrug therapy

“…22 Tumor treatment by ADEPT with alkylating agents, however, could prevent effective antitumor immunity due to dose-limiting leukopenia. 23,24 On the contrary, we found that rats cured by ADEPT with a prodrug of an alkylating agent developed potent antitumor immunity.…”