Study On Preparation And Anti-liver Cancer Effect Of Cryptotanshinone Inclusion Complex In Cyclodextrin

Objective: The low polarity and water solubility limit the clinical application of Cryptotanshinone.In this study,the cyclodextrin inclusion technique was used to improve the bioavailability of Cryptotanshinone.In addition,the anti-liver cancer activity of the inclusion compound was studied preliminarily,which may provide the basis for the new anticancer drugs of Cryptotanshinone.Methods: The extraction of total tanshinone was conducted according to Chinese Pharmacopoeia.Tanshinone IIA and Cryptotanshinone were isolated by the mean of silica gel column chromatography.The abilities of total tanshinone,Tanshinone IIA and Cryptotanshinone to inhibit the proliferation of human liver cancer Bel-7402 cells were compared by MTT assay.The inclusion compound(Cryptotanshinone-Hydroxylpropylbeta-cyclodextrin,CTan-HP-β-CD)was prepared by stirring-freeze-drying method.Using the entrapment rate and yield as the indexes,response surface methodology(RSM)was applied to optimize the preparation technology of CTan-HP-β-CD which was characterized by Infrared spectroscopy(IR)and Ultraviolet spectroscopy(UV).The mice with H22 tumor were established by subcutaneous-heterotopic transplantation.Use tumor-inhibition rate,thymus index and spleen index as indicators,the anti-liver cancer activity of CTan-HP-β-CD was preliminarily investigated in this study.Results: The extraction rate of total tanshinone was(1.15 ± 0.14)% and its purity measured by UV was(66.18 ± 1.56)%.The purity of Tanshinone IIA and Cryptotanshinone isolated by silica gel column chromatography were(59.23 ± 3.38)% and(64.54 ± 3.44)%,respectively.Cryptotanshinone had the stronger activity to inhibit Bel-7402 cells than Tanshinone IIA and total tanshinone,and its IC50 was 56.63 μg/m L.When mixture ratio was 6.04,temperature was 30 °C and reaction time was 3.1 h,preparation process of CTan-HP-β-CD was optimal,under this condition,the entrapment rate was(70.83 ± 0.15)%,yield was(92.68 ± 0.53)% and drug-loading rate was(7.08 ± 0.24)%.IR and UV demonstrated formation of inclusion.Animal experiments showed that the anti-liver cancer activity of CTan-HP-β-CD was significantly higher than that of Cryptotanshinone solution at the same dose.Compared with model group,CTan-HP-β-CD had no effect on thymus index and spleen index,while Cisplatin inhibited immune organs.Conclusion: Cyclodextrin inclusion technique could significantly improve the bioavailability of Cryptotanshinone.CTan-HP-β-CD could have anti-liver cancer activity in vitro and vivo.