Covalent binding studies on the 14C-labeled antitumor compound 2,5-bis(1-aziridinyl)-1,4-benzoquinone. Involvement of semiquinone radical in binding to DNA, and binding to proteins and bacterial macromolecules in situ.
2,5-Bis(1-aziridinyl)-1,4-benzoquinone (BABQ) is a compound from which several antitumour drugs are derived, such as Trenimone, Carboquone and Diaziquone (AZQ). The mechanism of DNA binding of BABQ was studied using 14C-labeled BABQ and is in agreement with reduction of the quinone moiety and protonation of the aziridine ring, followed by ring opening and alkylation. The one-electron reduced (semiquinone) form of BABQ alkylates DNA more efficiently than two-electron reduced or non reduced BABQ. Covalent binding to polynucleotides did not unambiguously reveal preference for binding to specific DNA bases. Attempts to elucidate further the molecular structure of DNA adducts by isolation of modified nucleosides from enzymatic digests of reacted DNA failed because of instability of the DNA adducts. The mechanism of covalent binding to protein (bovine serum albumin, BSA) appeared to be completely different from that of covalent binding to DNA. Binding of BABQ to BSA was not enhanced by reduction of the compound and was pH dependent in a way that is opposite to that of DNA alkylation. Glutathione inhibits binding of BABQ to BSA and forms adducts with BABQ in a similar pH dependence as the protein binding. The aziridine group therefore does not seem to be involved in the alkylation of BSA. Incubation of intact E. coli cells, which endogenously reduce BABQ, resulted in binding to both DNA and RNA, but also appreciable protein binding was observed.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Utrecht University, The Netherlands.
K J
KJ
Lusthof
N J
NJ
De Mol
L H
LH
Janssen
R J
RJ
Egberink
W
W
Verboom
D N
DN
Reinhoudt
D000478:Alkylation; D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism*; D001388:Aziridines; D001389:Azirines / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D016227:Benzoquinones / Q000378:metabolism*; D002417:Cattle; D003545:Cysteine / Q000494:pharmacology; D004247:DNA / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D010084:Oxidation-Reduction; D012329:RNA, Bacterial / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013997:Time Factors
D000970:Antineoplastic Agents; D001388:Aziridines; D001389:Azirines; D001426:Bacterial Proteins; D016227:Benzoquinones; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012710:Serum Albumin, Bovine; C025232:semiquinone radicals; C009970:ethylenimine quinone; D004247:DNA; D005978:Glutathione; D003545:Cysteine