title: Evidences for adduct formation between intracellular non-protein thiols and nitroazoles possessing an alpha,beta-unsaturated carbonyl side chain and the effects on radiosensitization of hypoxic cells.
abstract: Reactivity of a number of nitroazole derivatives bearing an alpha,beta-unsaturated carbonyl group on the side chain toward non-protein thiols (NPSH) was examined both in the phosphate buffer solution and in the biological system. These alpha,beta-unsaturated compounds reacted with NPSH, such as glutathione (GSH) and L-cysteine (Cys), in the buffer solution to afford the 1,4-addition products. The reaction gave a second-order rate constant. The adducts of methyl 4-(2'-nitroimidazol-1'-yl)crotonate (1) with GSH and Cys were isolated and characterized as two diastereomers (7a,b and 8a,b) in ca. 1:1 ratio, respectively. Similarly, exposure of EMT6/KU cells to 1 at 1.0 mM for 1 h resulted in depletion of the intracellular NPSH by more than 80%. Over 50% of the depleted NPSH was attributed to the formation of the conjugated diastereomeric adducts. On the other hand, incubation of EMT6/KU cells with 1 at 1.0 mM under hypoxic conditions before X-ray irradiation caused concurrently a sharp reduction of the shoulder of the dose-survival curves (reduced the extrapolation number (n) from 8.0 to ca. 1.0) and an increase in the slope (decreased the mean lethal dose (Do) to ca. 50% of the control level). The observed effects of 1 on the dose-survival curves were due to the NPSH depletion through the Michael addition occurred in the cellular system. A fairly linear relationship was obtained between the n value and the reduced intracellular NPSH level. It indicated that the shoulder effect of the dose-survival curves of hypoxic cells should be the result of the NPSH depletion by the alpha,beta-unsaturated carbonyl group attached to the nitroazoles.
authors.affiliation: Department of Energy and Hydrocarbon Chemistry, Graduate School of Engineering, Kyoto University, Japan. lingz@scl.kyoto-u.ac.jp
authors.forename: L
authors.initials: L
authors.lastname: Zhou
authors.forename: A
authors.initials: A
authors.lastname: Ino
authors.forename: W M
authors.initials: WM
authors.lastname: Dai
authors.forename: S
authors.initials: S
authors.lastname: Nishimoto
mesh_terms: D001393:Azoles / Q000737:chemistry* / Q000494:pharmacology; D002021:Buffers; D015687:Cell Hypoxia; D002478:Cells, Cultured; D003545:Cysteine / Q000737:chemistry; D004307:Dose-Response Relationship, Radiation; D005978:Glutathione / Q000737:chemistry; D006801:Humans; D010710:Phosphates / Q000502:physiology; D011836:Radiation Tolerance / Q000187:drug effects*; D013438:Sulfhydryl Compounds / Q000737:chemistry* / Q000494:pharmacology
chemical_list: D001393:Azoles; D002021:Buffers; D010710:Phosphates; D013438:Sulfhydryl Compounds; D005978:Glutathione; D003545:Cysteine