Photodynamic activity of monocationic and non-charged methoxyphenylporphyrin derivatives in homogeneous and biological media

Abstract

A novel 5-[4-(trimethylammonium)phenyl]-10,15,20-tris(2,4,6-trimethoxyphenyl)porphyrin iodide (2) has been synthesized. A positive charge was incorporated at a peripheral position to increase the amphiphilic character of the structure. The photodynamic effect of the cationic porphyrin 2 was compared with that of non-charged 5-(4-aminophenyl)-10,15,20-tris(2,4,6-trimethoxyphenyl)porphyrin (1), both in a homogeneous medium bearing photooxidizable substrates and in vitro on the Hep-2 human larynx carcinoma cell line. Absorption and fluorescence spectroscopic studies in different media show that 2 is essentially unaggregated in solution, and also in human cells. The singlet molecular oxygen, O₂(¹Δ_g), production was evaluated using 9,10-dimethylanthracene in N,N-dimethylformamide, yielding Φ_Δ values of ∼0.66 for both porphyrins. The addition of β-carotene suppresses the O₂(¹Δ_g)-mediated photooxidation. L-Tryptophan and guanosine 5′-monophosphate were used as biological substrate models. Porphyrin 2 sensitizes the decomposition of both compounds faster than does 1. In the biological medium, no dark cytotoxicity was observed, even though a high porphyrin concentration (10 µM) and a long incubation time (24 h) were employed. Cell treatments were performed with 5 µM of porphyrin for 24 h. Under these conditions, the uptake of porphyrin 2 into Hep-2 was about 3 times higher than that of 1. Cell survival after irradiation with visible light was dependent upon both the light exposure level and intracellular sensitizer concentration. Thus, a higher photocytotoxic effect was found for porphyrin 2 in comparison to 1. These results show that the amphiphilic monocationic porphyrin 2 could be a promising model for phototherapeutic agents with potential applications in tumor cell inactivation by photodynamic therapy.