title: A transient product of DNA alkylation can be stabilized by binding localization.
abstract: A 9-aminoacridine conjugate of a silyl-protected bis(acetoxymethyl)phenol (bisQMP) was synthesized and evaluated as an inducible cross-linking agent of DNA to test our ability to harness the chemistry of reactive quinone methide intermediates (QM). The acridine component was chosen for its ability to delivery an appendage to the major groove of DNA, and the silyl-protected component was chosen for its ability to generate two quinone methide equivalents in tandem upon addition of fluoride. This design created competition between reaction of (1) the 2-amino group of guanine that reacts irreversibly to form a stable QM adduct and (2) the more nucleophilic N7 group of guanine that reacts more efficiently but reversibly to form a labile QM adduct. This lability was apparently compensated by co-localization of the N7 group and QM in the major groove since the N7 adduct appeared to dominate the profile of products formed by duplex DNA. The controlling influence of acridine was also expressed in the sensitivity of the conjugate to ionic strength. High salt concentration inhibited covalent reaction just as it inhibits intercalation of the cationic acridine. As expected for QM formation, the presence of fluoride was indeed necessary for initiating reaction, and no direct benzylic substitution was observed. The conjugate also cross-linked DNA with high efficiency, forming one cross-link for every four alkylation events. Both alkylation and cross-linking products formed by duplex DNA were labile to hot piperidine treatment which led to approximately 40% strand scission and approximately 50% reversion to a material with an electrophoretic mobility equivalent to the parent DNA. All guanines exhibited at least some reactivity including those which were recalcitrant to cross-linking by an oligonucleotide-bisQMP conjugate designed for triplex formation [Zhou, G.; Pande, P.; Johnson, A. E.; Rokita, S. E. Bioorg. Med. Chem. 2001, 9, 2347-2354].
authors: Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA.
authors: Willem F
authors: WF
authors: Veldhuyzen
authors: Praveen
authors: P
authors: Pande
authors: Steven E
authors: SE
authors: Rokita
mesh_terms: D000477:Alkylating Agents / Q000138:chemical synthesis / Q000737:chemistry*; D000478:Alkylation; D000585:Aminacrine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D045563:Indolequinones / Q000737:chemistry; D007700:Kinetics; D009994:Osmolar Concentration; D010636:Phenols / Q000138:chemical synthesis / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D000477:Alkylating Agents; D003432:Cross-Linking Reagents; D045563:Indolequinones; D010636:Phenols; C068040:quinone methide; D000585:Aminacrine; D004247:DNA