Dynamics of Oxygen-Independent Photocleavage of Blebbistatin as a One-Photon Blue or Two-Photon Near-Infrared Light-Gated Hydroxyl Radical Photocage

Development of versatile, chemically tunable photocages for photoactivated chemotherapy (PACT) represents an excellent opportunity to address the technical drawbacks of conventional photodynamic therapy (PDT) whose oxygen-dependent nature renders it inadequate in certain therapy contexts such as hypoxic tumors. As an alternative to PDT, oxygen free mechanisms to generate cytotoxic reactive oxygen species (ROS) by visible light cleavable photocages are in demand. Here, we report the detailed mechanisms by which the small molecule blebbistatin acts as a one-photon blue light-gated or two-photon near-infrared light-gated photocage to directly release a hydroxyl radical (•OH) in the absence of oxygen. By using femtosecond transient absorption spectroscopy and chemoselective ROS fluorescent probes, we analyze the dynamics and fate of blebbistatin during photolysis under blue light. Water-dependent photochemistry reveals a critical process of water-assisted protonation and excited state intramolecular proton transfer (ESIPT) that drives the formation of short-lived intermediates, which surprisingly culminates in the release of •OH but not superoxide or singlet oxygen from blebbistatin. CASPT2//CASSCF calculations confirm that hydrogen bonding between water and blebbistatin underpins this process. We further determine that blue light enables blebbistatin to induce mitochondria-dependent apoptosis, an attribute conducive to PACT development. Our work demonstrates blebbistatin as a controllable photocage for •OH generation and provides insight into the potential development of novel PACT agents

Ultrafast Time-Resolved Spectroscopic Study on the Photophysical and Photochemical Reaction Mechanisms of <i>ortho</i>-Methylbenzophenone in Selected Solutions

The photophysical and photochemical reaction pathways of ortho-methylbenzophenone (o-MeBP) in different solutions were investigated by employing femtosecond to nanosecond transient absorption and nanosecond time-resolved resonance Raman spectroscopy methods. In pure acetonitrile, neutral or pH 1 aqueous solutions, o-MeBP exhibit similar excited-state evolutions upon excitation in which o-MeBP will experience excitation to an excited state then undergo intersystem crossing and solvent arrangement followed by 1,5 hydrogen atom transfer processes to form the first singlet excited state, triplet state (n, π*), biradical intermediates, and enol form transients, respectively. However, in a pH 0 acidic solution, the protonation of o-MeBP will form the cation biradical intermediate that facilitates radical coupling to generate a benzocyclobutanol product, which causes a dramatic reduction of the lifetime of the enol form transients. In contrast, in sodium bicarbonate solution, the biradical intermediate may be quenched by the bicarbonate ion to construct a C–C bond and form the carboxylic acid that causes a fast decay of biradical intermediate. These results demonstrate that the photophysical and photochemical reaction pathways of o-MeBP are pH-dependent in aqueous solution which may be very useful for the capture of CO2 capture by photoexcitation of aromatic ketones