Photodynamic inhibition of acetylcholinesterase after two-photon excitation of copper tetrasulfophthalocyanine.

Sequential two-photon (2-gamma) activated copper tetrasulfophthalocyanine (CuPcS(4)) was shown capable of inactivating acetylcholinesterase (ACE). ACE activity was measured photometrically by the Ellman method. Simultaneous irradiation of ACE in the presence of CuPcS(4) with 514 nm (183 mW/cm(2)) and 670 nm (86 mW/cm(2)) continuous wave (CW) light induced a 20-50% increase in enzyme inhibition as compared to one-photon (1-gamma) irradiation, using either 514- or 670-nm (CW) light at the same fluences. The enzyme activity was not affected by CuPcS(4) or light alone, decreased linearly with the irradiation time, and was shown to be oxygen-dependent. We conclude that the photoinactivation of ACE with sequential 2-gamma irradiation involves reactive oxygen species produced by the interaction of the upper excited T(n) state of CuPcS(4) with molecular oxygen. As CuPcS(4) shows little activity as a conventional 1-gamma photosensitizer, unwanted side effects such as prolonged skin sensitivity are eliminated rendering 2-gamma photodynamic therapy advantageous for the treatment of selected medical applications.