In vivo mutagenesis induced by CC-1065 and adozelesin DNA alkylation in a transgenic mouse model.
Although considerable work has focused on characterizing the bonding chemistry and sequence selective alkylation of DNA by cyclopropylpyrroloindole compounds, little is known about the molecular consequence of their N-3-adenine adducts in whole animal systems. We have utilized a transgenic mouse system, harboring a lambda phage shuttle vector, to assess the mutagenic potential of the antitumor compounds CC-1065 and adozelesin and, for the first time, to track the in vivo fate of their unique DNA modifications at the nucleotide level. Mice were inoculated with a single therapeutic dose of these agents and sacrificed at either 18 h, 3 days, or 15 days for extraction and analysis of liver DNA. Mutant frequencies obtained from drug treated and control animals were determined by in vitro packaging of the phage vector from genomic DNA followed by a colorimetric plaque assay to screen for phage in which the accompanying lacI repressor gene had mutated. Although undetectable at 18 h posttreatment, by 72 h a 3-fold increase in mutant frequency was observed in drug treated animals such that sequence analysis of drug induced mutations could be performed and a direct comparison made between in vitro and in vivo DNA alkylation. Base substitution involving guanine or cytosine accounted for 64% of the 41 mutations sequenced from drug treated animals. Only 7 of the mutations occurred at a cyclopropylpyrroloindole alkylation site while 23 occurred 1 to 4 nucleotides from a potentially alkylated adenine.
Monroe
T J
TJ
Medicinal Chemistry Research, Upjohn Company, Kalamazoo, Michigan 49001.
Mitchell
M A
MA
D000478:Alkylation; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D000970:Antineoplastic Agents / Q000633:toxicity*; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D001572:Benzofurans; D003509:Cyclohexanecarboxylic Acids / Q000378:metabolism / Q000633:toxicity*; D053138:Cyclohexenes; D004247:DNA / Q000378:metabolism*; D000080890:Duocarmycins; D007211:Indoles; D007933:Leucomycins / Q000378:metabolism / Q000633:toxicity*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D016296:Mutagenesis
D000903:Antibiotics, Antineoplastic; D000970:Antineoplastic Agents; D001572:Benzofurans; D003509:Cyclohexanecarboxylic Acids; D053138:Cyclohexenes; D000080890:Duocarmycins; D007211:Indoles; D007933:Leucomycins; C018782:CC 1065; C060090:adozelesin; D004247:DNA