aluminum-phthalocyanine-disulfonate and aluminum-tetrasulfophthalocyanine

Photodynamic therapy is an attractive technique for various skin tumors and non-cancerous skin lesions. However, while the aim of photodynamic therapy is to target and damage only the malignant cells, it unavoidably affects some of the healthy cells surrounding the tumor as well. However, data on the effects of PDT to normal cells are scarce, and the characterization of the pathways activated after the photodamage of normal cells may help to improve clinical photodynamic therapy. In our study, primary human epidermal keratinocytes were used to evaluate photodynamic treatment effects of photosensitizers with different subcellular localization. We compared the response of keratinocytes to lysosomal photodamage induced by phthalocyanines, aluminum phthalocyanine disulfonate (AlPcS

Topics: Apoptosis; Autophagy; Cell Differentiation; Computer Simulation; Humans; Indoles; Isoindoles; Keratinocytes; Kinetics; Lysosomes; Mesoporphyrins; Models, Biological; Organometallic Compounds; Photochemotherapy; Photosensitizing Agents

The sulfonated phthalimidomethyl phthalocyanine hydroxyl aluminum (Al(OH)PcSP) is an amphiphilic photosensitizer which was proved to have the photodynamic activities against cancer. The electronic absorption spectra and aggregation state of the Al(OH)PcSP in aqueous alcoholic solutions and water were investigated. The results showed that the Al(OH)PcSP existed in the form of monomer in aqueous alcoholic solutions. The increase in the carbon chain and hydroxy of alcohol in solutions had no significant effects on the absorption spectra behaviors of Al(OH)PcSP. But in water, the Al(OH)PcSP existed in the equilibrium between monomer and dimer. The dimerization constant was 5.7307 x 10(4) mol-1.L. The characteristic absorption peak of the Q band of the Al(OH)PcSP dimer was red shifted to 740.5 nm from that of the monomer (676.5 nm), which contrast with those of other metal phthalocyanines dimer. The study on the fluorescence spectra of Al(OH)PcSP in aqueous alcoholic solutions suggested that the fluorescence of the dimer was weak.

Topics: Electron Probe Microanalysis; Indoles; Kinetics; Organometallic Compounds; Photosensitizing Agents; Phthalimides; Solutions; Spectrometry, Fluorescence; Spectrophotometry, Atomic; Structure-Activity Relationship; Sulfonic Acids

Nude mice were given AlPcS2a (aluminum phthalocyanine disulfonate) and AlPcS4 (aluminum phthalocyanine tetrasulfonate) by intraperitoneal injections. After time intervals of 1-48 hours the mice were exposed to 150 mW cm-2 light at 670 nm and the phthalocyanine fluorescence was measured during light exposure. During the first few minutes of light exposure the phthalocyanine fluorescence of the skin of the mice increased by up to a factor of two, indicating lysosomal localization of the dye and permeabilization of the lysosomes. The process did not occur in the skin of dead mice, indicating that the process was dependent on oxygen.

Topics: Aluminum; Animals; Female; Indoles; Light; Mice; Mice, Inbred BALB C; Mice, Nude; Organometallic Compounds; Radiation-Sensitizing Agents; Spectrometry, Fluorescence

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.

Topics: Animals; Female; Hematoporphyrin Derivative; Indoles; Mammary Neoplasms, Experimental; Mice; Mice, Inbred Strains; Neoplasm Transplantation; Organometallic Compounds; Photochemotherapy; Radiation-Sensitizing Agents; Skin; Tissue Distribution

Important differences exist in the responses to photodynamic agents of normal and tumour-derived pancreatic acinar cells. In the present study amylase release has been used to assess the mechanisms by which the photodynamic drugs tetra- and disulphonated aluminium phthalocyanine (A1PcS4, A1PcS2) act on pancreatic cells via energy and calcium-dependent activation and transduction pathways. The photodynamic release of amylase was found to be energy dependent and inhibited by the chelation of free cytoplasmic calcium but not by the removal of extracellular calcium. In contrast to their effects on normal acinar cells, the photodynamic action of A1PcS4 and A1PcS2 was to inhibit amylase secretion from pancreatoma AR4-2J cells. Removal of extracellular calcium reversed this inhibitory effect on AR4-2J cells and produced a significant increase in amylase release, but chelation of free cytoplasmic calcium did not affect the inhibitory photodynamic action of the phthalocyanines on amylase release from the tumour cells. Overall, these results demonstrate further important distinctions between the photodynamic action of sulphonated aluminium phthalocyanines on normal versus tumour exocrine cells of the pancreas and indicate that calcium plays an important role in photodynamic drug action, since these agents affected intracellular calcium mobilisation at some distal point in the membrane signal transduction pathway for regulated secretion. Furthermore, the photodynamic inhibition of constitutive secretion in tumour cells may involve a calcium-dependent membrane target site or modulation of membrane calcium channels by activation of protein kinase C.

Topics: Amylases; Animals; Antimycin A; Bethanechol; Calcium; Carcinoma, Acinar Cell; Deoxyglucose; Egtazic Acid; Indoles; Male; Oligomycins; Organometallic Compounds; Pancreas; Pancreatic Neoplasms; Photochemotherapy; Radiation-Sensitizing Agents; Rats; Rats, Sprague-Dawley; Signal Transduction; Sincalide; Stimulation, Chemical; Tumor Cells, Cultured

V79 cells incubated with di- or tetrasulfonated aluminium phthalocyanines (AlPcS2 or AlPcS4) showed a granular fluorescence pattern. Co-staining with the lysosomotropic dye acridine orange (AO) indicated that the granules that were stained by these photoactive phthalocyanines were identical to lysosomes. Small light exposures made the lysosomes permeable to the dyes without inactivating the cells. Also, the lysosomal enzymes beta-AGA and cathepsin (L+B) were inactivated by small light exposures when AlPcS4 was present. Such small and almost nontoxic light exposures caused a redistribution of the dyes in the cells that was accompanied by a more than 10-fold increase in the fluorescence quantum yields of the dyes. Surprisingly, this redistribution and increase in fluorescence did not result in any significant increase in the photosensitivity of the cells.

Topics: Acetylglucosaminidase; Acridine Orange; Animals; Cathepsin B; Cathepsin L; Cathepsins; Cells, Cultured; Cricetinae; Cricetulus; Cysteine Endopeptidases; Endopeptidases; Fibroblasts; Fluorescent Dyes; Indoles; Light; Lysosomes; Organometallic Compounds; Photochemotherapy; Photosensitizing Agents; Spectrometry, Fluorescence

Levels of disulfonated and tetrasulfonated aluminum phthalocyanines (AlPcS2,4) were measured in cells derived from FsaR tumors (murine fibrosarcoma) using a fluorescence-activated cell sorter (FACS). The tumors were excised from animals injected with the sensitizer 24 h earlier and enzymatically dissociated. Before flow cytometry, the cells were stained with fluorescein isothiocyanate-conjugated anti-mouse monoclonal antibodies to specific immune cell membrane markers (Mac1, Fc receptor (FcR) or CD45). Staining to FcR and CD45 was combined with a DNA stain Hoechst 33342. This enabled concomitant discrimination to be made by the FACS between different populations of tumor-infiltrating host cells and malignant cells. The results showed on average 1.49 times higher AlPcS2 levels and 1.16 times higher AlPcS4 levels in Mac1-positive (Mac1+) compared with Mac1-negative (Mac1-) tumor cell populations. The same type of experiments performed with SCCVII tumor (squamous cell carcinoma) gave average Mac1+/Mac1- ratios of 1.75 and 1.45 for AlPcS2 and AlPcS4 respectively. The data using other antibodies and DNA staining are consistent with the conclusion that, based on average per cell content, elevated levels of AlPcS2, and to a lesser extent AlPcS4, are retained in tumor-associated macrophages (TAM). The levels of these photosensitizers in other leukocytes and in non-immune host cells were not substantially different from those in malignant tumor cells. It is also shown that elevated levels of AlPcS2 and AlPcS4 are not localized in all TAM, but rather in a fraction of this cell population characterized by extremely high photosensitizer content.

Topics: Animals; Benzimidazoles; Biomarkers; Female; Fibrosarcoma; Flow Cytometry; Fluorescein-5-isothiocyanate; Indoles; Macrophage-1 Antigen; Mice; Mice, Inbred C3H; Organometallic Compounds; Radiation-Sensitizing Agents; Sarcoma, Experimental; Staining and Labeling; Tissue Distribution

By means of laser scanning fluorescence microscopy the intratumoral localization patterns of several photosensitizers in LOX tumors in nude mice were studied. Lipophilic dyes such as TPPS1 (tetraphenylporphine monosulfonate), TPPS2a (tetraphenylporphine disulfonates with the sulfonate groups on adjacent rings), AlPCS1 (aluminium phthalocyanine monosulfonate) and AlPCS2 (aluminium phthalocyanine disulfonates) localized mainly in tumor cells. The fluorescence intensity of these dyes increased from 4 h to 48 h postinjection and the fluorescence was still observable 120 h postinjection. The more hydrophilic dyes such as TPPS3 (tetraphenylporphine trisulfonates), TPPS4 (tetraphenylporphine tetrasulfonates), and AlPCS4 (aluminium phthalocyanine tetrasulfonates) localized mainly extracellularly in the tumorous stroma. The fluorescence intensity of these dyes decreased from 4 h to 48 h postinjection. 120 h postinjection no significant fluorescence of these dyes could be seen in the tumors. P-II (Photofrin II), 3-THPP [tetra(3-hydroxyphenyl)porphine], TPPS2o (tetraphenylporphine disulfonates with the sulfonate groups on opposite rings) and AlPCS3 (aluminum phthalocyanine trisulfonates) had a combined localization pattern, i.e. a strongly cytoplasmic membrane-localizing pattern and a weakly intracellular distribution pattern, although some fluorescence could be seen in the tumorous stroma. The data are discussed in relation to what is known about the in vivo photosensitizing efficiency of some of the dyes.

Topics: Animals; Cell Line; Dihematoporphyrin Ether; Female; Fluorescent Dyes; Hematoporphyrins; Humans; Indoles; Lasers; Melanoma; Mice; Mice, Inbred BALB C; Mice, Nude; Microscopy, Phase-Contrast; Organometallic Compounds; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents

We have previously reported photodynamic therapy of normal rat colon using aluminium sulphonated phthalocyanine (AISPc). In that study, the AISPc used was a mixture of phthalocyanines of different degrees of sulphonation. Phthalocyanines of defined degrees of sulphonation have recently become available and we compared the distribution of the di- and tetra-sulphonates (AIS2Pc and AIS4Pc) in rat colon and colon wall structures employing both chemical extraction and fluorescence photometry using a charge coupled device imaging system. Also, the photodynamic effects produced by these components in rat colon were compared at various times after photosensitization. After intravenous photosensitizer administration using equimolar doses, the concentration of AIS2Pc in colon fell off more rapidly with time than AIS4Pc. Differences were noted in the microscopic distribution of these compounds, with the di-sulphonate exhibiting peak fluorescence in colon wall structures by 1 h after photosensitization, while mucosal fluorescence with the tetra-sulphonate peaked at 5 h. Fluorescence was also lost from the colon wall much more slowly with the tetra-sulphonate, which tended to be retained in the submucosa. Maximum photosensitizing capability was seen at 1 h with AIS2Pc and no lesions could be produced with photodynamic therapy at 1 week, with up to 5.65 mumol/kg. With AIS4Pc (5.65 mumol/kg), while no lesions could be produced with light treatment at 1 h, photodynamic therapy at 1 week produced lesions only slightly smaller than those produced with treatment at 48 h (the time of maximum effect), and significant photosensitization was present at 2 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Aluminum; Animals; Colon; Female; Indoles; Necrosis; Organometallic Compounds; Photochemotherapy; Radiation-Sensitizing Agents; Rats; Rats, Inbred Strains