Photophysics and light induced photobiology of antiviral and antitumor agents, hypericin and hypocrellin
Is Version Of
The dissertation mainly focuses on the excited-state photophysics and the light-induced biological activity of hypericin and its analogs.;Femtosecond laser technology has provided the opportunity to investigate the rapid dynamics of these molecules. Fluorescence upconversion measurements, which monitor only emission from the fluorescent singlet state, demonstrate that hexamethoxy hypericin, which possesses no labile protons, has an instantaneous rise time for its transient response. On the other hand, hypericin shows a clear 10-ps rise time. This confirms excited-state H-atom transfer as the primary photophysical process in hypericin.;Femtosecond transient absorption spectroscopy technique is used to determine if excited state H-atom transfer is concerted. Previous studies using human serum albumin (HSA) and hypericin suggested that excited state H-atom transfer is concerted, but the results from the hypericin in reverse micelles show no evidence for a concerted hydrogen atom transfer mechanism. We are, however, unable to conclude if only one hydrogen atom is transferred or if two are transferred in a stepwise fashion.;By means of time-resolved infrared spectroscopy, ab initio quantum mechanical calculations, and synthetic organic chemistry, a region in the infrared spectrum, between 1400 and 1500 cm-1, of triplet hypericin has been found corresponding to translocation of the hydrogen atom between the enol and the keto oxygens, O &cdots; H &cdots; O. This result is discussed in the context of the photophysics of hypericin and of eventual measurements to observe directly the excited-state H-atom transfer.;Light-induced antiviral activity of hypericin and hypocrellin is compared in normoxic and hypoxic conditions. Although both molecules require oxygen to show full virucidal effects, hypericin is still effective at low oxygen level where hypocrellin is not. Since the singlet oxygen yield of hypericin is about half of that of hypocrellin, this result cannot be explained by a traditional Type II mechanism. We propose that the ejected proton upon illumination might enhance the activity of activated oxygen species.;A series of hypericin analogs were found to differ in their cytotoxic activity induced by ambient light levels. These analogs vary in their ability to partition into cells, to generate singlet oxygen as well as in other photophysical properties. The percent distribution of hypericin and its analogs in cells are measured using a steady state absorption technique. We attempt to find a relationship between those results and the exact localization of the drug at subcellular level.