Potent Antioxidant and Anti-Tyrosinase Activity of Butein and Homobutein Probed by Molecular Kinetic and Mechanistic Studies

Butein (BU) and homobutein (HB) are bioactive polyhydroxylated chalcones widespread in dietary plants, whose antioxidant properties require mechanistic definition. They were investigated by inhibited autoxidation kinetic studies of methyl linoleate in Triton (TM) X-100 micelles at pH 7.4, 37 degrees C. Butein had k(inh) = (3.0 +/- 0.9) x 10(4) M(-1)s(-1) showing a chain-breaking mechanism with higher antioxidant activity than reference alpha-tocopherol (k(inh) = (2.2 +/- 0.6) x 10(4) M(-1)s(-1)), particularly concerning the stoichiometry or peroxyl radical trapping n = 3.7 +/- 1.1 vs. 2.0 for tocopherol. Homobutein had k(inh) = (2.8 +/- 0.9) x 10(3) M(-1)s(-1), pairing the relative BDEOH measured by radical equilibration EPR as 78.4 +/- 0.2 kcal/mol for BU and estimated as 82.6 kcal/mol for HB. The inhibition of mushroom tyrosinase (mTYR) by HB and BU was also investigated. BU gives a reversible uncompetitive inhibition of monophenolase reaction with K-I ' = 9.95 +/- 2.69 mu M and mixed-type diphenolase inhibition with K-I = 3.30 +/- 0.75 mu M and K-I ' = 18.75 +/- 5.15 mu M, while HB was nearly competitive toward both mono- and diphenolase with respective K-I of 2.76 +/- 0.70 mu M and 2.50 +/- 1.56 mu M. IC50 values (monophenolase/diphenolase at 1 mM substrate) were 10.88 +/- 2.19 mu M/15.20 +/- 1.25 mu M, 14.78 +/- 1.05 mu M/12.36 +/- 2.00 mu M, and 33.14 +/- 5.03 mu M/18.27 +/- 3.42 mu M, respectively, for BU, HB, and reference kojic acid. Molecular docking studies confirmed the mechanism. Results indicate very potent antioxidant activity for BU and potent anti-tyrosinase activity for both chalcones, which is discussed in relation to bioactivity toward protection from skin disorders and food oxidative spoilage.