Formaldehyde-activated pixantrone is a monofunctional DNA alkylator that binds selectively to CpG and CpA doublets

Benny J Evison, Francis Chi Keung Chiu, Gabriella Pezzoni, Don R Phillips, Suzanne M Cutts

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26 Citations (Scopus)

Abstract

The topoisomerase II poison mitoxantrone is important in the clinical management of human malignancies. Pixantrone, a novel aza-anthracenedione developed to improve the therapeutic profile of mitoxantrone, can efficiently alkylate DNA after formaldehyde activation. In vitro transcriptional analysis has now established that formaldehyde-activated pixantrone generates covalent adducts selectively at discrete CpG and CpA dinucleotides, suggesting that the activated complex binds to guanin or cytosine (or both) bases. The stability of pixantrone adduct-induced transcriptional blockages varied considerbaly, reflecting a mixture of distinct pixantrone adduct types that may include relatively labile monoadducts and more stable interstrand cross-links. 6,9-Bis-[[2-(dimethylamino)ethyl]amino]benzo[g]isoquinoline-5,10-dione (BBR 2378), the dimethyl N-substituted analog of pixantrone, could not form adducts, suggesting that pixantrone alkylates DNA through the primary amino functions located in each side chain of the drug. Pixantrone generated DNA adducts only when guanin was present in substrates and exhibited a lack of adduct formation with inosine-containing poly-nucleotides, confirming that the N2 amino group of guanine is the site for covalent attachment of the drug. Mass spectrpmetric analysis of oligonucleotide-drug complexes confimred that formation of covalent pixantrone-DNA adducts is mediated by a single methylene linkage provided by formaldehyde and that this occurs only with guanine-containing double stranded oligonucletide substrates. CpG methylation, an epigenetic modification of the mammalian genome, significantly enhanced the generation of pixantrone-DNA adducts within a methylated DNA substrate, indicating that the methylated dinucleotide may be a favoured target in a cellular environment.
Original languageEnglish
Pages (from-to)184 - 194
Number of pages11
JournalMolecular Pharmacology
Volume74
Issue number1
DOIs
Publication statusPublished - 2008

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