New reaction pathways of dopamine under oxidative stress conditions: Nonenzymatic iron-assisted conversion to norepinephrine and the neurotoxins 6-hydroxydopamine and 6,7-dihydroxytetrahydroisoquinoline

Aerial oxidation of dopamine at concentrations as low as 50 mu M in the pre sence of ferrous ions in phosphate buffer (pH 7.4) led in the early stages (6-8 h) to the formation of the quinone of the neurotoxin B-hydroxydopamine , 2, followed (24 h) by a complex product pattern comprising main component s norepinephrine (5), 3,4-dihydroxybenzaldehyde (4), and the neurotoxic alk aloid 6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (3). Product formation r equired the assistance of metal ions such as Mn(II), Zn(II), and iron, in e ither the ferrous or ferric form. Product yields were shown to vary linearl y with iron and dopamine concentration in the early phases of the reaction (2 h). Biologically relevant antioxidants, like glutathione and ascorbate, and metal chelators, e.g., 2,2'-bipyridyl, inhibited dopamine conversion to products 2-5, but not substrate consumption, while hydroxyl radical scaven gers such as DMSO and mannitol did not alter the course of the reaction. On the contrary, mannitol increased product yields, an effect seen for other monosaccharides. Catalase exhibited a significant inhibitory effect particu larly on the formation of 3 and 4. By using O-18(2), evidence was obtained for incorporation of the label into the carbonyl oxygen of 4, but not into the hydroxyl group of 5. On the basis of these and other results, a complet e mechanistic picture of the oxidation is drawn involving conversion of dop amine to the corresponding o-quinone and its quinonemethide tautomer with c oncomitant reduction of Oa to H2O2 Nucleophilic attack by H2O to the quinon emethide gives rise to 5, while H2O2 addition leads to benzaldehyde 4 via a beta-amino-hydroperoxide intermediate. This latter reaction path also give s formaldehyde which yields the isoquinoline 3 by Pictet-Spengler condensat ion with dopamine. The quinone 2 results from H2O2 attack at the 6-position of dopamine o-quinone in agreement with previous studies. These results pr ovide an insight into new routes of nonenzymatic conversion of dopamine to its metabolite norepinephrine and neurotoxic species which may become opera tive under conditions relevant to neurodegeneration.