Reductive activation of diaziquone and possible involvement of free radicals and the hydroquinone dianion.
To analyze the mode of action of diaziquone [AZQ] on DNA, we examined the activity of two AZQ analogues and N,N',N"-triethylenethiophosphoramide on three forms [supercoiled (Form I), open circular (Form II), and linear (Form III)] of PM-2 DNA. The AZQ analogues contained chlorine atoms which substituted either the carbethoxyamino groups or the aziridine groups of the parent compound. N,N',N"-triethylenethiophosphoramide is a triaziridine compound containing pentavalent phosphorus instead of a quinone group. We found that only when reduced with sodium borohydride did AZQ change the topology of the three forms of PM-2 DNA by introducing mainly single strand breaks. The AZQ analogue containing only aziridines (RQ2) was active in both its oxidized and its reduced forms, while the analogue containing only the carbethoxyamino groups (RQ14) or N,N',N"-triethylenethiophosphoramide were not active in either form. Under similar experimental conditions, Adriamycin alone altered the electrophoretic mobility of PM-2 DNA, while borohydride reduced Adriamycin did not. By using electron spin resonance spectroscopy, we showed that dihydrodiaziquone (AZQH2) oxidizes to the semiquinone in the presence of oxygen. Although AZQH2 was active against DNA, it was not active against cellular DNA synthesis as measured by [3H]thymidine incorporation into exponentially growing HEp-2 cells. However, AZQ alone prevented [3H]thymidine incorporation into HEp-2 cells. We found that HEp-2 cells have the ability to reduce AZQ to its free radical anion, but AZQH2 does not autoxidize to the free radical in the presence of cells. The reductive ability of HEp-2 cells may be responsible in part for preventing the oxidation of AZQH2 to the free radical. We found that under our conditions (1-h incubations) the aziridines are essential for the activity of aziridinyl quinones against PM-2 DNA and that in the case of AZQ the hydroquinone is also required.
D001388:Aziridines / Q000494:pharmacology; D001389:Azirines; D001435:Bacteriophages; D016227:Benzoquinones; D002460:Cell Line; D004249:DNA Damage; D004261:DNA Replication / Q000187:drug effects; D004278:DNA, Superhelical / Q000187:drug effects; D004279:DNA, Viral / Q000187:drug effects; D004317:Doxorubicin / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals; D006801:Humans; D006873:Hydroquinones; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D011809:Quinones / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013852:Thiotepa / Q000494:pharmacology
D001388:Aziridines; D001389:Azirines; D016227:Benzoquinones; D004278:DNA, Superhelical; D004279:DNA, Viral; D005609:Free Radicals; D006873:Hydroquinones; D011809:Quinones; C043462:NSC 191295; C043461:NSC 30705; D004317:Doxorubicin; D013852:Thiotepa; C029108:diaziquone; C031927:hydroquinone