In Vitro Inhibition of Xanthine Oxidase Purified from Arthritis Serum Patients by Nanocurcumin and Artemisinin Active Compounds

Abstract

Curcumin and artemisinin are commonly used in traditional East Asian medicine. In this study, we investigated the inhibitory effects of these active compounds on xanthine oxidase (XO) using allopurinol as a control. XO was purified from the serum of arthritis patients through ammonium sulfate precipitation (65%) and ion exchange chromatography on diethylaminoethyl (DEAE)-cellulose. The specific activity of the purified enzyme was 32.5 U/mg protein, resulting in a 7-fold purification with a yield of 66.8%. Molecular docking analysis revealed that curcumin had the strongest interaction energy with XO, with a binding energy of -9.28 kcal/mol. The amino acid residues Thr1077, Gln762, Phe914, Ala1078, Val1011, Glu1194, and Ala1079 were located closer to the binding site of curcumin than artemisinin, which had a binding energy of -7.2 kcal/mol. In vitro inhibition assays were performed using nanocurcumin and artemisinin at concentrations of 5, 10, 15, 20, and 25 µg/mL. Curcumin inhibited enzyme activity by 67-91%, while artemisinin had a lower inhibition ratio, which ranged from 40-70% compared to allopurinol as a control.

Keywords: artemisinin; curcumin; inhibition; molecular docking; xanthine oxidase.

Scheme 1

Chemical structure of (a) curcumin and (b) artemisinin.

Figure 1

Different views of the nanocurcumin of scales, (a) 150 nm and (b) 100 nm.

Figure 2

XRD test for the nanocurcumin.

Figure 3

The DEAE-cellulose column chromatography (3 × 25 cm) of Xanthine oxidase purified from arthritis patients’ sera, which was eluted with 0.05 M potassium phosphate buffer at pH 7.5 using a 0.1–1 M NaCl gradient concentration at a flow rate of 30 mL/h.

Figure 4

Simulation of binding nanocurcumin with Xanthine Oxidase: (a) 3D and (b) 2D views.

Figure 4

Simulation of binding nanocurcumin with Xanthine Oxidase: (a) 3D and (b) 2D views.