Studies On The Hepatoprotective Activity Of Kinsenoside And The Synthesis Of Its Fully Acetylated Derivative

Anoectochilus roxburghii (Wall.) Lindl is an orchidaceous perennial herbaceous plant. Anoectochilus is precious traditional Chinese medicinal materials, widely used as a folk medicine for diabetes, hypertension, liver diseases, diuretic, cancer and other diseases, without any toxic or side effects. The main ingredients of Anoectochilus are glycoside, alkaloid, sterides, flavone, taurine, amino acids, trace elements and so on. The results from the present investigation suggest that Anoectochilus directly act on the liver cell membrane, then protect integrity of the liver cell membrane and prevent harmful substances damage to it, and prevent the pathological changes to necrosis. The survival natural environment of Anoectochilus has been greatly changed. Because of the sharp reduction in wild resources and the low rate of natural reproduction, coupled with the scarcity of natural resource, now is endangered. Anoectochilus is widely use in the folk medicine and it is effective, but its natural resources is scarce. So we research on novel total synthesis fully acetylated derivative of kinsenoside under the guidance of the hepatoprotective activity. The research methods and results of the article.1. Study on activity of kinsenoside and the product of fully acetylated of kinsenoside.The hepatoprotective effect of kinsenoside and the product of fully acetylated of kinsenoside were tested on CCl4 induced liver injury in mice. Intraperitoneal injection of 0.125% CCl4 oil solution was carried out to establish the model, to study hepatoprotective effect on different dose of two different compounds in liver injury and effect of ALT and AST index value in mice serum. The results show that ALT and AST index value of the two groups of compounds in different doses were both lower than model group. Moreover, it was clearly dose-dependent. Histopathological examination are also show that the effect of kinsenoside and fully acetylated product of kinsenoside in various doses on hepatocyte swelling, necrosis and inflammatory cell infiltration are significant improved compare with the model group. The product of fully acetylated of kinsenoside show the same hepatoprotectiv activity as silymarin, but more than that of Anoectochilus glycosides. 2. Synthesis of the fully acetylated of kinsenoside.Our experimental group has been working on natural medicine research to search for active ingredients. By studying on the chemical composition of Anoectochilus, we found kinsenoside as the active ingredients. In order to get adequate samples for pharmacological activity and structure reconstruction, we conducted synthesis of the compound.The product in theory can be divided into two parts: aglycone and glycon. For the aglycone of (S)-3-hydroxy-γ-butyrolactone we use lactose and maltodextrin as the starting material through three different ways to synthesis. Finally, we have chosen method 3 as the most preferred synthesis method, which uses maltodextrin as raw material, cumene hydroxyperoxide as catalyst, and microwave irradiation involved in the response through One-Pot Synthesis of (S)-3-hydroxy-γ-butyrolactone. The part of glycon we synthesis two intermediates are 2,3,4,6-O-Tetraacetyl-α-D-glucopyranosyl bromide and 2,3,4,6-O-Tetraacetyl-α-D-glucopyranosyl trichloroacetimidate. We synthesis 2,3,4,6-O-Tetraacetyl-α-D-glucopyranosyl trichloroacetimidate through two different ways , and Method 2 is better which improves the reaction yield and purity.For target compound, we also carried out two different ways to explore. For the reaction of glycosides, we also use 2,3,4,6-O-Tetraacetyl-α-D- glucopyranosyl bromide and 2,3,4,6-O-Tetraacetyl-α-D-glucopyranosyl trichloroacetimidate for intermediates. Finally, we chose line 2 to synthesis of target compound, which uses TMSOTf as catalyst and 2,3,4,6-O-Tetraacetyl-α-D- glucopyranosyl trichloroacetimidate as intermediates of glycon. The physical and chemical properties and spectral data of target compound are consistent with the literature. It provide basis for study of kinsenoside structure reconstruction.