Abstract
A chemical extraction assay and fluorescence microscopy incorporating a light-sensitive thermoelectrically cooled charge-coupled device (CCD) camera was used to study the kinetics of uptake, retention and localisation of disulphonated aluminium phthalocyanine (A1PcS2) and tetrasulphonated aluminium phthalocyanine (A1PcS4) at different time intervals after an i.p. injection at a dose of 10 mg kg-1 body weight (b.w.) in tumour and surrounding normal skin and muscle of female C3D2/F1 mice bearing CaD2 mammary carcinoma. Moreover, the photodynamic effect on the tumour and normal skin using sulphonated aluminium phthalocyanines (A1PcS1, A1PcS2, A1pcS4) and Photofrin was compared with respect to dye, dye dose and time interval between dye administration and light exposure. The maximal concentrations of A1PcS2 in the tumour tissue were reached 2-24 h after injection of the dye, while the amounts of A1PcS4 peaked 1-2 h after the dye administration. A1PcS2 was simultaneously localised in the interstitium and in the neoplastic cells of the tumour, whereas A1PcS4 appeared to localise only in the stroma of the tumour. The photodynamic efficiency (light was applied 24 h after dye injection at a dose of 10 mg kg-1 b.w.) of the tumours was found to decrease in the following order: A1PcS2 > A1PcS4 > Photofrin > A1PcS1. Furthermore, photodynamic efficacy was strongly dependent upon dye doses and time intervals between dye administration and light exposure: the higher the dose, the higher the photodynamic efficiency. The most efficient photodynamic therapy (PDT) of the tumour was reached (day 20 tumour-free) when light exposure took place 2 h after injection of A1PcS2 (10 mg kg-1). A dual intratumoral localisation pattern of the dye, as found for A1PcS2, seems desirable to obtain a high photodynamic efficiency. The kinetic patterns of uptake, retention and localisation of A1PcS2 and A1PcS4 are roughly correlated with their photodynamic effect on the tumour and normal skin.
Highlights
(b.Ax in tumour and surrounding normal skin and muscle of female C-D F, mice bearing CaD2 mammary carcinoma
Observations that phthalocyamines had affinity for tumour tissues were documented more than 30 years ago. interest in phthalocvanines as second-generation photosensitisers for photodynamic therapy (PDT) of cancer arose in 1985 w-hen Ben-Hur and Rosenthal (1985) reported that some phthalocyanines were efficient photosensitisers in mammalian cells
At present. most studies on Pcs related to PDT have been conducted with water-soluble sulphonated M-Pcs. in particular sulphonated aluminium phthalocvanines (AlPcS,s)
Summary
Derivatives of aluminium phthalocyanines with mono-. diand tetrasulphonate groups Diand tetrasulphonate groups (AlPcSj. AlPcS, and AlPcS,). These derivatives were assessed by high-performance liquid chromatography (HPLC) to be >90% pure The dye called AlPcS, in the present study probably contains two sulphonate groups on adjacent phenyl rings (AlPcS2,). Stock solutions of AlPcS2 and AIPcS4 were prepared in Dulbecco's phosphate-buffered saline (PBS) (Gibco), while AlPcS, was dissolved initially in a small amount of 40% ethanol in PBS followed by dilution in PBS. All solutions of AlPcS,s were sonicated for 5 min (Elma Transsonic, type T400, Germany) before use in order to reduce the degree of aggregation. All chemicals used were of the highest purity commercially available
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