Cytotoxic ROS-Consuming Mn(III) Synzymes: Structural Influence on Their Mechanism of Action

ROS (i.e., reactive oxygen species) scavenging is a key function of various Mn-based enzymes, including superoxide dismutases (SODs) and catalases, which are actively linked to oxidative stress-related diseases. In this study, we synthesized and characterized two novel Mn(III)-based synzymes (i.e., synthetic enzymes), designated <b>C1</b> ([Mn<b>L1</b>Cl(H₂O)]Cl·3H₂O) and <b>C2</b> ([Mn<b>L2</b>Cl₂]·2H₂O), which differ in the presence of a bridging aliphatic or aromatic group in the chelator. Using a range of analytical techniques, we found that the aromatic <b>C2</b> bridge significantly influences the Mn(III) center's <i>cis-β</i> configuration, unlike <b>C1</b>, which adopts a <i>trans</i> configuration. We then thoroughly evaluated the oxidation-reduction properties of <b>C1</b> and <b>C2</b>, including their redox potentials (by cyclic voltammetry) and capacity to consume various ROS species (using DPPH, hydroxyl radical, hydrogen peroxide, and superoxide UV-visible spectrophotometric assays). The specific kinetics of the H₂O₂ dismutation process, as measured by a Clark-type electrode and time-resolved ESI-MS, revealed that both synzymes possess catalytic activity. Toxicological experiments using the <i>Galleria mellonella</i> larval model demonstrated the compounds' innocuous nature towards higher eukaryotic organisms, while cytotoxicity assays confirmed their selective efficacy against lung cancer cells. Additional cytological assays, such as the thiobarbituric acid reactive substances assay and caspase-3 activity and p53 expression analysis, reported that <b>C1</b> and <b>C2</b> induce cytotoxicity against cancer cells via apoptosis rather than necrosis and behave very differently towards redox substances and ROS-regulating enzymes in vivo. These findings suggest that the structural differences between <b>C1</b> and <b>C2</b> lead to distinct redox properties and biological activities, highlighting the potential of these novel Mn(III)-based synzymes as therapeutic agents for the treatment of oxidative stress-related diseases, particularly lung cancer. Further studies are warranted to elucidate the underlying mechanisms of action and explore their clinical applications.