title: Hypoxia-selective, enzymatic conversion of 6-nitroquinoline into a fluorescent helicene: pyrido[3,2-f]quinolino[6,5-c]cinnoline 3-oxide.
abstract: Regions of low oxygen concentration (hypoxia) occur in both normal human physiology and under pathophysiological conditions. Fluorescent probes for the direct imaging of cellular hypoxia could be useful tools that complement radiochemical imaging and immunohistochemical staining methods. In this work, we set out to characterize the hypoxia-selective enzymatic metabolism of a simple nitroaryl probe, 6-nitroquinoline (1). We envisioned that this compound might undergo hypoxia-selective, bioreductive conversion to the fluorescent product, 6-aminoquinoline (2). The probe 1 was, indeed, converted to a fluorescent product selectively under hypoxic conditions by the one-electron reducing enzyme NADPH:cytochrome P450 reductase. However, inspection of the fluorescence spectrum and LC-MS analysis of the reaction mixture revealed that the expected product 2 was not formed. Rather, the 63-fold increase in fluorescence emission at 445 nm resulting from the hypoxic metabolism of 1 was due to formation of the azoxy-helicene product, pyrido[3,2-f]quinolino[6,5-c]cinnoline 3-oxide (4). The generation of 4 involves an unusual biaryl bond formation under reductive conditions. The mechanism of this process remains uncertain but could proceed via combination of a nitroaryl radical anion with a neutral nitrosoaryl radical, followed by tautomerization and intramolecular condensation between the resulting hydroxylamine and nitroso functional groups. Bioreductive metabolism of nitroaryl compounds represents a promising strategy for the selective delivery of cytotoxic agents and fluorescent markers to hypoxic tissue, but the results described here provide an important glimpse of the chemical complexity that can be associated with the enzymatic one-electron reduction of nitroaryl compounds.
authors.affiliation: Department of Chemistry, University of Missouri, 125 Chemistry Building, Columbia, Missouri 65211, USA.
authors.forename: Anuruddha
authors.initials: A
authors.lastname: Rajapakse
authors.forename: Kent S
authors.initials: KS
authors.lastname: Gates
mesh_terms: D000634:Aminoquinolines / Q000737:chemistry*; D015687:Cell Hypoxia; D004583:Electrons; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D000860:Hypoxia / Q000139:chemically induced*; D015394:Molecular Structure; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry* / Q000378:metabolism; D009596:Nitrophenols / Q000737:chemistry*; D010084:Oxidation-Reduction; D011083:Polycyclic Compounds / Q000737:chemistry*; D011725:Pyridines / Q000737:chemistry*; D011804:Quinolines / Q000737:chemistry*
chemical_list: D000634:Aminoquinolines; D005456:Fluorescent Dyes; D009596:Nitrophenols; D011083:Polycyclic Compounds; D011725:Pyridines; D011804:Quinolines; C031660:helicenes; C574414:pyrido(3,2-f)quinolino(6,5-c)cinnoline 3-oxide; C032928:6-aminoquinoline; D009251:NADPH-Ferrihemoprotein Reductase