3-7-dihydro-6-(4-(2-(n--(5-fluoresceinyl)thioureido)ethoxy)phenyl)-2-methylimidazo-(1-2-a)pyrazin-3-one and Disease-Models--Animal

A novel method of photodynamic diagnosis of cancer mediated by chemiluminescence probe is presented. The mechanism for photodynamic therapy involves singlet oxygen ((1)O(2)) generated by energy transfer from photosensitizers. (1)O(2) can react with 3,7-dihydro-6-[4-[2-(N'-(5-fluoresceinyl)thioureido)ethoxy]phenyl]-2-methylimidazo[1,2-a]pyrazin-3-one sodium salt (FCLA), which is a Cypridina luciferin analog and a specific chemiluminescence probe for detecting (1)O(2) and superoxide (O(2)(-)). The reaction of FCLA and (1)O(2) can give emission with peak wavelength at about 532 nm. In the present study, FCLA was chosen as an optical reporter of (1)O(2) produced from the photosensitization reaction of hematoporphyrin derivative in model solution and in nude mice with transplanted mammary cancer. Photosensitized chemiluminescence from the reaction of FCLA with (1)O(2) was detected by a highly sensitive Intensified Charge-Coupled Device detector. The chemiluminescence was markedly inhibited by the addition of 10 mmol/l sodium azide (NaN(3)) to the model solution and minor effects were observed at the addition of 10 micromol/l superoxide dismutase, 20 mmol/l mannitol and 100 microg/ml catalase, respectively, thus indicating that (1)O(2) generation from photosensitization reaction mainly results in light emission. Experiments in vivo with tumor-bearing mice showed a clear chemiluminescence image of tumor. The study suggests that this novel method may be applicable to the diagnosis of superficial tumors.

Topics: Animals; Diagnostic Techniques and Procedures; Disease Models, Animal; Female; Fluoresceins; Hematoporphyrins; Imidazoles; Luminescent Measurements; Mice; Models, Chemical; Molecular Probes; Neoplasms; Photosensitizing Agents; Singlet Oxygen; Sodium Azide

As a chemiluminescence (CL) probe, 3,7-dihydro-6-[4-[2-(N'-(5-fluoresceinyl) thioureido)ethoxy]phenyl]-2-methylimidazo[1,2-a]pyrazin-3-one dosium salt (FCLA) can sensitively and specifically react with singlet oxygen ((1)O(2)) and superoxide (O(2)(-)). Based on the fact that photosensitization action involves (1)O(2) generation, this report presents a novel method for localization of tumor mediated by FCLA. In experiments, FCLA was used as an optical reporter of (1)O(2) produced from photosensitization reaction of hematoporphyrin derivative. The effects of azide inhibition and deuterium enhancement on photosensitized CL indicated that (1)O(2) molecules generated during photosensitization action could be measured and imaged. In vivo experiments in vivo with tumor-bearing nude mice showed a clear CL image of tumor. The study suggests that CL imaging method may be applicable to the detection of (1)O(2) and the diagnosis of superficial tumors.

Topics: Animals; Disease Models, Animal; Fluoresceins; Humans; Imidazoles; Luminescent Measurements; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Middle Aged; Neoplasms, Experimental; Photosensitizing Agents; Reactive Oxygen Species; Singlet Oxygen; Sodium Azide