New anti-hepatocellular injury drugs with better curative effects and fewer side effects are urgently needed at present. In this study, a series of novel <i>N</i>-acetylcysteine (NAC) derivatives were designed, synthesized and biologically evaluated for their anti-hepatocellular injury activities against two different cell models. In the biological evaluation against hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>)-induced LO2 hepatocytes, half of the target compounds exhibited moderate to potent activities in improving the model cell viability, and two compounds (<b>6a</b> and <b>6b</b>) displayed more potent activities in decreasing malondialdehyde (MDA) levels than the positive control NAC. In further 4-acetamidophenol (APAP)-induced LO2 cell experiment, compounds <b>6a</b> and <b>6b</b> could not only improve the cell viability but also significantly reduce the secretion of MDA. Additionally, compound <b>6a</b> displayed excellent Caco-2 permeability and oral bioavailability in rats. All these experimental results suggested that compounds <b>6a</b> and <b>6b</b> could serve as potential lead molecules for further development of anti-hepatocellular injury drugs.