Scavenging effects of metal complexes of water-soluble thiacalix[4]arenetetrasulfonate on superoxide anion radicals

The scavenging effects of metal complexes of thiacalix4]arenetetrasulfonate (Me-TCAS4], Me=H?, Fe3(+), Mn3(+), Mn2(+), Cu2(+), and Zn2(+)) on superoxide anion radicals (O??) generated from the xanthine-xanthine oxidase system were investigated by the nitroblue tetrazolium (NBT) method and electron spin resonance (ESR) spin-trapping method using 5,5-dimethyl-1-pyrroline-N-oxide as a trapping reagent. As a reference, calix4]arenetetrasulfonate (H?)-CAS4]), calix6]arenehexasulfonate (H?-CAS6]) and calix8]areneoctasulfonate (H?-CAS8]) were also examined. The results by the NBT method indicated that Fe3(+)- and Mn3(+)-TCAS4] exhibited the highest O?? scavenging activity among Me-TCAS4] and H?-CASn] (n = 4, 6, 8) in this study. The IC?? values of Fe3(+)- and Mn3(+)-TCAS4] for O?? scavenging activity were estimated to be 5.3 and 7.8 μM, respectively, and were almost the same as those of tannin acid, catechin and their derivatives, which are known as very effective scavengers of O??. Scavenging activities were in the order of Fe3(+)- and Mn3(+)-TCAS4]>Mn2(+)-, Cu2(+)-, and Zn(2+)-TCAS4]>H(2)-TCAS4] and H?-CASn] (n=4, 6, 8). Each activity of Me-TCAS4] (Me=Fe3(+), Mn3(+), Mn2(+), Cu2(+), and Zn2(+)) was higher than that of the corresponding metal ion, indicating that H?-TCAS4] has the ability to raise the activity of the metal ion itself by forming a complex. Also, the ESR spin-trapping method revealed that Fe3(+)- and Mn3(+)-TCAS4] showed high O?? scavenging activities, similarly to the results by the NBT method.