Primary liver cancer is the sixth most commonly diagnosed cancer worldwide and the third leading cause of cancer deaths in 2020 [1]. Hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC) are the two main types of primary liver cancer [2], [3]. Among them, HCC accounts for more than 80 % of primary liver cancers [4], [5]. Currently, the main treatments for HCC are surgical resection, transplantation, ablation, transarterial chemoembolization and targeted therapies with sorafenib and Lenvatinib [6], [7]. Despite continuous advances in treatment, HCC remains poor prognostic, and effective treatment is difficult for most patients [8], [9]. Therefore, it is urgent to develop a new method for HCC treatment with little side effects.

Over recent years, natural products have received more and more attention due to their superior efficacy and decreased toxicity [10] E. tomentosus is a traditional folk medicine for the treatment of a variety of diseases, mainly in southern China [11]. Germacrolides are important sesquiterpene lactone of E. tomentosus [12]. Previous studies have shown that germacrolides have various pharmacological activities, including antitumor, anti-inflammatory, and so on [13], [14]. The highly oxidized germacrolides contain one oxygen bridge that divides the ten-membered ring into a 5/8 ring system [12]. Although the cytotoxic activity of germacrolides-based compounds has been well studied, the specific mechanism is still unclear.

In this study, 9 highly oxidized germacrolides (WBDC-1, WBDC-2, WBDC-3, WBDC-4, WBDC-5, WBDC-6, WBDC-7, WBDC-8, WBDC-9) isolated from E. tomentosus in our previous study [12] were screened for their inhibitory activity against HCC cells, showing that WBDC-1 exhibited better inhibitory activity against HCC cells. In this study, the anti-HCC effect of WBDC-1 was evaluated in vitro and in vivo. Network pharmacology was used to find the targets of compounds and a variety of biological methods were used to verify.