Cycloimide bacteriochlorin p derivatives: photodynamic properties and cellular and tissue distribution
Résumé
Reactive oxygen species generated by photosensitizers are efficacious remedy for tumor eradication. Eleven cycloimide derivatives of bacteriochlorin p (CIBCs) with different N-substituents at the fused imide ring and various substituents replacing the 3-acetyl group were evaluated as photosensitizers with special emphasis on structure–activity relationships. The studied CIBCs absorb light within a tissue transparency window (780–830 nm) and possess high photostability at prolonged light irradiation. The most active derivatives are 300-fold more phototoxic toward HeLa and A549 cells than the clinically used photosensitizer Photogem due to the substituents that improve intracellular accumulation (distribution ratio of 8–13) and provide efficient photoinduced singlet oxygen generation (quantum yields of 0.54–0.57). The substituents predefine selective CIBC targeting to lipid droplets, Golgi apparatus, and lysosomes or provide mixed lipid droplets and Golgi apparatus localization in cancer cells. Lipid droplets and Golgi apparatus are critically sensitive to photoinduced damage. The average lethal dose of CIBC-generated singlet oxygen per volume unit of cell was estimated to be 0.22 mM. Confocal fluorescence analysis of tissue sections of tumor-bearing mice revealed the features of tissue distribution of selected CIBCs and, in particular, their ability to accumulate in tumor nodules and surrounding connective tissues. Considering the short-range action of singlet oxygen, these properties of CIBCs are prerequisite to efficient antitumor photodynamic therapy. Abbreviations: AO, acridine orange; BODIPY ceramide, N-((4-(4,4-difluoro-5-(2-thienyl)-4-bora-3a, 4a-diaza-s-indacene-3-yl) phenoxy)acetyl)sphingosine; CIBC(s), cycloimide derivative(s) of bacteriochlorin p; CIC(s), cycloimide derivative(s) of chlorin p6; CrEL, polyoxyethylene derivative of hydrogenated castor oil, Cremophor EL; CSI, confocal spectral imaging; DR, distribution ratio, a ratio of average cytoplasmic concentration to the extracellular concentration of a photosensitizer; FCS, fetal calf serum; LSCM, laser scanning confocal microscopy; MTT, 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide; NIR, near infrared; PDT, photodynamic therapy; ΦΔ, singlet oxygen quantum yield; RNO, 4-nitroso-N,N,-dimethylaniline; Rh123, rhodamine 123; ROS, reactive oxygen species