Viola G, Facciolo L, Dall'Acqua S, Di Lisa F, Canton M, Vedaldi D, Fravolini A, Tabarrini O, Cecchetti V
Department of Pharmaceutical Sciences, University of Padova, via Marzolo 5, 35131 Padova, Italy.
Toxicol In Vitro. 2004 Oct;18(5):581-92. doi: 10.1016/j.tiv.2004.01.008.
Three selected aminoquinolones endowed with a potent antibacterial (compounds 1 and 2) and antiviral activity (compound 3) have been evaluated for their phototoxic properties in vitro. Photostability studies of these compounds indicate that compound 3 is photostable whereas compound 1 and in particular, compound 2 are rapidly photodegraded upon UVA irradiation, yielding a toxic photoproduct. Intracellular localization of these compounds has been evaluated by means of fluorescence microscopy using tetramethylrhodamine methyl ester and acridine orange, which are specific fluorescent probes for mitochondria and lysosomes, respectively. No co-staining was observed with lysosomal stain for all the test compounds. On the contrary compound 3 was found to be specifically incorporated in mitochondria. The compounds exhibited remarkable phototoxicity in two cell culture lines: human promyelocytic leukaemia (HL-60) and human fibrosarcoma (HT-1080). The quinolone-induced photodamage was also evaluated measuring the photosensitizing cross-linking in erythrocyte ghost membranes, the strand breaks activity and oxidative damage on plasmid DNA. The results show that these derivatives are able to photoinduce crosslink of erythrocytes spectrin, whereas do not significantly photocleavage DNA directly, but single strand breaks were observed after treatment of photosensitized DNA with two base excision repair enzymes, Fpg and Endo III respectively.
已对三种具有强效抗菌活性(化合物1和2)和抗病毒活性(化合物3)的氨基喹诺酮类化合物进行了体外光毒性特性评估。这些化合物的光稳定性研究表明,化合物3具有光稳定性,而化合物1,尤其是化合物2在UVA照射下会迅速光降解,产生有毒的光产物。已分别使用四甲基罗丹明甲酯和吖啶橙通过荧光显微镜评估了这些化合物的细胞内定位,这两种分别是线粒体和溶酶体的特异性荧光探针。所有测试化合物均未观察到与溶酶体染色的共染色。相反,发现化合物3特异性地掺入线粒体中。这些化合物在两种细胞系中表现出显著的光毒性:人早幼粒细胞白血病(HL-60)和人纤维肉瘤(HT-1080)。还通过测量红细胞血影膜中的光敏交联、链断裂活性以及对质粒DNA的氧化损伤来评估喹诺酮诱导的光损伤。结果表明,这些衍生物能够光诱导红细胞血影蛋白交联,而不会直接显著光切割DNA,但在用两种碱基切除修复酶Fpg和Endo III分别处理光敏DNA后观察到单链断裂。