Effect of β-cyclodextrin encapsulation on cytotoxic activity of acetylshikonin against HCT-116 and MDA-MB-231 cancer cell lines

Acetylshikonin (AcSh), as a red colored pigment found in roots of the plants from family Boraginaceae, showed excellent cytotoxic activity. Due to its hydrophobic nature, and thus poor bioavailability, the aim of this study was to prepare acetylshikonin/beta-cyclodextrin (AcSh/(beta-CD) inclusion complex by using coprecipitation method, characterize obtained system by using UV/VIS, IR and H-1 NMR spectroscopy, and determine cytotoxic activity. Phase solubility test indicated formation of A(L)-type binary system (substrate/ligand ratio was 1:1 M/M), with stability constant Ks of 306.01 M-1. Formation of noncovalent bonds between inner layer of the hole of beta-CD and AcSh was observed using spectroscopic methods. Notable changes in chemical shifts of two protons (-0.020 ppm) from naphthoquinone moiety (C-6-H and C-7-H), as well as protons from hydroxyl groups (-0.013 and -0.009, respectively) attached to C-5 and C-8 carbons from naphthoquinone part indicate that the molecule of AcSh enters the beta-CD cavity from the aromatic side. Cytotoxic activity against HCT-116 and MDA-MB-231 cell lines was measured by MTT test and clonogenic assay. Mechanisms of action of free AcSh and inclusion complex were assessed by flow cytometry. In comparison to free AcSh, AcSh/beta-CD showed stronger short-term effect on HCT-116 cells and superior long-term effect on both cell lines. Inclusion complex induced more pronounced cell cycle arrest and autophagy inhibition, and induced increase in accumulation of intracellular ROS more effectively than free AcSh. In conclusion, AcSh/beta-CD binary system showed better performances regarding cytotoxic activity against tested tumor cell lines. (C) 2019 The Author(s). Published by Elsevier B.V. on behalf of King Saud University.