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Research Article | Open Access

5-Demethylnobiletin and its major metabolites: efficient preparation and mechanism of their anti-proliferation activity in HepG2 cells

Yanping Xina,b,c,1, Ting Zhengc,1, Man Zhangc, Ruiqiang Zhanga, Siyue Zhuc, Dongli Lia,b(), Denggao Zhaoa,b, Yanyan Maa,b, Chi-Tang Hoc, Qingrong Huangc()
School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, China
International Healthcare Innovation Institute (Jiangmen), Jiangmen 529020, China
Department of Food Science, Rutgers University, New Jersey 08901, USA

1 These two authors contribute equally.

Peer review under responsibility of KeAi Communications Co., Ltd.

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Abstract

5-Demethylnobiletin (5-DMN), a hydroxylated polymethoxyflavone (OH-PMF) identified in aged citrus peels, has demonstrated health benefiting effects in previous studies. 5-DMN undergoes biotransformation in vivo, yielding 5,3'-didemethylnobiletin (5,3'-DDMN), 5,4'-didemethylnobiletin (5,4'-DDMN) and 5,3',4'-tridemethylnobiletin (5,3',4'-TDMN). However, the anti-cancer effects of 5-DMN and its in vivo metabolites against HepG2 cells remain unclear. In this study, an efficient chemical synthetic method was developed to obtain 5-DMN and its 3 metabolites, and their molecular structures were confirmed by 1H NMR and LC-MS. Cytotoxicity, cell cycle arrestment, apoptosis and caspase-3 expression were investigated to evaluate the anti-liver cancer effects of these OH-PMFs on HepG2 cells. The results showed that all 4 compounds inhibited the proliferation of HepG2 cells in a concentration-dependent manner. Their anti-proliferative activity was exerted through inducing G2/M phase arrestment, cell apoptosis and promoting expression of a key apoptotic protein called cleaved caspase-3. Our results indicated that 5,3'-DDMN and 5,3',4'-TDMN showed a stronger inhibitory activity on cell proliferation than 5-DMN, followed by 5,4'-DDMN. The expression of cleaved caspase-3 was the highest in cells treated with 5,4'-DDMN, implying that the apoptosis induced by other OH-PMFs might be mediated by other apoptotic execution proteins. Our research reveals the application potential and scientific evidence for the production and functionality of OH-PMFs.

Keywords

Hydroxylated polymethoxyflavonesChemical synthesisHepG2Cell cycleApoptosis

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Food Science and Human Wellness
Volume 11 Issue 5,
September 2022
Pages 1191-1200
DOI: 10.1016/j.fshw.2022.04.018
Cite this article:
Xin Y, Zheng T, Zhang M, et al. 5-Demethylnobiletin and its major metabolites: efficient preparation and mechanism of their anti-proliferation activity in HepG2 cells. Food Science and Human Wellness, 2022, 11(5): 1191-1200. https://doi.org/10.1016/j.fshw.2022.04.018
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