Acute oral toxicity of damnacanthal and its anticancer activity against colorectal tumorigenesis

Warunya Woradulayapinij; Apipu Pothiluk; Thararat Nualsanit; Thunyatorn Yimsoo; Werayut Yingmema; Pleumchitt Rojanapanthu; Yukyung Hong; Seung Joon Baek; Worapapar Treesuppharat

doi:10.1016/j.toxrep.2022.10.015

Acute oral toxicity of damnacanthal and its anticancer activity against colorectal tumorigenesis

Toxicol Rep. 2022 Oct 29:9:1968-1976. doi: 10.1016/j.toxrep.2022.10.015. eCollection 2022.

Authors

Warunya Woradulayapinij¹, Apipu Pothiluk¹, Thararat Nualsanit², Thunyatorn Yimsoo³, Werayut Yingmema³, Pleumchitt Rojanapanthu⁴, Yukyung Hong⁵, Seung Joon Baek⁵, Worapapar Treesuppharat¹

Affiliations

¹ Thammasat University Research Unit in Mechanisms of Drug Action and Molecular Imaging, Drug Discovery and Development Center, Office of Advanced Science and Technology, Thammasat University, Pathum Thani 12120, Thailand.
² Chulabhorn International College of Medicine, Thammasat University, Pathum Thani 12120, Thailand.
³ Laboratory Animal Center, Office of Advanced Science and Technology, Thammasat University, Pathum Thani 12120, Thailand.
⁴ Drug Discovery and Development Center, Office of Advanced Science and Technology, Thammasat University, Pathum Thani 12120, Thailand.
⁵ Laboratory of Signal Transduction, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 08826, South Korea.

Abstract

Damnacanthal is an anthraquinone, extracted, and purified from the root of Morinda citrifolia in Thailand. This study aimed to measure acute oral toxicity and to investigate the anticancer activity of damnacanthal in colorectal tumorigenesis. We found that the growth of human colorectal cancer cells was inhibited by damnacanthal in a dose- and a time-dependent manner. The growth inhibitory effect of damnacanthal was better than that of 5-FU used as a positive control in colorectal cancer cells, along with the downregulation of cell cycle protein cyclin D1. Similarly, an oral treatment of damnacanthal effectively inhibited the growth of colorectal tumor xenografts in nude mice, which was approximately 2-3-fold higher as compared to 5-FU by tumor size as well as expression of bioluminescence. Furthermore, the study of acute oral toxicity in mice exhibited a relatively low toxicity of damnacanthal with a LD₅₀ cut-off value of 2500 mg/kg according to OECD Guideline 423. These results reveal the potential therapeutic activity of a natural damnacanthal compound as an anti-colorectal cancer drug.

Keywords: 5-FU, 5-Fluorouracil; ALT, Alanine aminotransferase; AST, Aspartate aminotransferase; Acute oral toxicity; Anticancer activity; BSA, Bovine serum albumin; BUN, Blood urea nitrogen; Colorectal tumorigenesis; D20, Damnacanthal at 20 mg/kg; D40, Damnacanthal at 40 mg/kg; DMSO, Dimethyl sulfoxide; DPBS, Dulbecco’s phosphate buffered saline; Damnacanthal; F20, 5-Fluorouracil at 20 mg/kg; FBS, Fetal bovine serum; FTIR, Fourier transform infrared spectroscopy; IC50, Half-maximal inhibitory concentration; LD50, Median lethal dose; MS, Mass spectrometry; MTT, 3-(4,5-Dimethythiazol-2-yl)− 2,5-diphenyltetrazolium bromide; NC, Negative control; NMR, Nuclear magnetic resonance spectroscopy; PMSF, Phenylmethanesulfonyl fluoride; TBST, Tris-buffered saline containing 0.05 % Tween 20; TLC, Thin layer chromatography; VLC, Vacuum liquid chromatographic method.