Assessment of the hepatoprotective effects of Bicyclol's forced degradation products using a zebrafish model.

Bicyclol (BIC), a synthetic hepatoprotective agent widely prescribed in China, lacks comprehensive safety and activity profiles for its degradation products (DPs). Here, we systematically investigated BIC's forced degradation behavior using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Three hydrolytic degradation products (DP1-DP3) were isolated via HPLC and structurally characterized, revealing methylenedioxy group hydrolysis as the primary degradation pathway. Molecular docking simulations demonstrated enhanced target binding affinities of DPs compared to BIC, supported by absorption, distribution, metabolism, and excretion (ADME) predictions showing improved drug-likeness. In an alcohol-induced fatty liver zebrafish model, both BIC and its DPs attenuated hepatic macrovesicular steatosis and inflammatory responses. Mechanistically, treatment normalized lipid metabolism by downregulating alcohol-induced expression of FASN, SREBP1, PPARα, and PPARγ, while reduced IL-6 and TNF-α levels confirmed anti-inflammatory efficacy. These findings demonstrate that BIC DPs exhibit dual pharmacological activity through lipid homeostasis modulation and inflammation suppression, providing critical insights for quality control and therapeutic applications.