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Objectives: The main purpose of this study was to enhance the intestinal absorption activity and hepatoprotective effect of herpetrione by drug nanosuspensions.<br>Methods: Herpetrione nanosuspensions (HNS) were prepared using pH-dependent dissolving-precipitating/homogenization process and then systematically characterized. The intestinal absorption activity of HNS were studied using the recirculating perfusion technique in comparison with herpetrione coarse suspensions (HCS) and pure herpetrione using the recirculating perfusion technique. The protective effect of HNS against acute liver injury induced by carbon tetrachloride (CCl₄) in mice was also investigated and compared with that of HCS.<br>Key findings: The mean particle size of HNS was 269 ± 7 nm with a polydispersity index of 0.187 ± 0.021. The result of X-ray powder diffraction indicated that herpetrione was in amorphous state in both coarse powder and nanosuspensions. The intestinal absorption activity of HNS were superior to the HCS and pure herpetrione. As evidenced by the lowering of serum aminotransferase levels and the improvement of the degree of liver lesion, pretreatment with HNS markedly enhanced the hepatoprotective effect of herpetrione against acute liver injury induced by CCl₄ in mice.<br>Conclusion: HNS prepared using pH-dependent dissolving-precipitating/homogenization technique are able to significantly enhance the intestinal absorption activity and the hepatoprotective effect of herpetrione due to the particle size reduction.