title: DNA Sequence Preference and Adduct Orientation of Pyrrolo[2,1-c][1,4]benzodiazepine Antitumor Agents.
abstract: The pyrrolobenzodiazepines (PBDs) are covalent DNA minor-groove binding agents with a reported preference for binding to 5'-Pu-G-Pu sequences with their A rings oriented toward the 3'-end of the covalently modified DNA strand. Using HPLC/MS methodology and a range of designed hairpin-forming 17-mer oligonucleotides, the kinetics of reaction of a bis-pyrrole PBD conjugate (GWL-78, 2) has been evaluated with eight isomeric oligonucleotides, each containing a single PBD binding site in one of two locations. The PBD-binding base pair triplets were designed to include every possible combination of A and T bases adjacent to the covalently reacting guanine. Contrary to expectations, 2 reacted most rapidly with TGT and TGA sequences, and adducts were observed to form in both the 3'- and the 5'-directions. Molecular modeling studies revealed that for 3'-oriented adducts, this preference could be explained by formation of a hydrogen bond between the N10-H of the PBD and the oxygen of the C2-carbonyl of a thymine base on the 3'-side of the covalently bound guanine. For 5'-adducts, an analogous PBD N10-H hydrogen bond may form instead to the N3 of an equivalent adenine on the opposite strand.
authors.affiliation: Gene Targeting Drug Design Research Group.
authors.forename: Khondaker M
authors.initials: KM
authors.lastname: Rahman
authors.affiliation: Gene Targeting Drug Design Research Group.
authors.forename: Higia
authors.initials: H
authors.lastname: Vassoler
authors.affiliation: Gene Targeting Drug Design Research Group.
authors.forename: Colin H
authors.initials: CH
authors.lastname: James
authors.affiliation: Gene Targeting Drug Design Research Group ; Spirogen Ltd.
authors.forename: David E
authors.initials: DE
authors.lastname: Thurston
keywords: DNA; GWL-78; Pyrrolo[2,1-c][1,4]benzodiazepine (PBD); antitumor agents; minor-groove binder