Chemical Composition Study Of High Polar Fraction Of TCM Rhizoma Heterosmilacis Japonicae

Objective : Baituling(Rhizoma Heterosmilacis Japonicae) is the dried rhizome of Heterosmilax yunnanensis Gagnep., Heterosmilax japonica Kunth and Heterosmilax chinensis Wang which belong to the genus Heterosmilax Kunth of family Liliaceae. They were mainly distributed in the province of Shaanxi, Gansu, Anhui, Sichuan, Yunnan and other places. Due to the effects of improving the mobility of joints,removing damp, counteracting toxicity and clearing heat, they had been widely used for the treatment of morbid leukorrhea, dysuria with turbid discharge, chronic eczema, contracture of limbs and muscle pain in syphilis etc. On the other hand, Baituling was the composition of compound Kushen injection(CKI) which is widely used on the clinic as anti-tumor medicine. The previous pharmacological experiment showed that Baituling can effectively inhibit the growth of tumor, especially the water-soluble part, so the systematically studies on its secondary metabolite and quality control were carried out.Methods:Secondary metabolites from Baituling(Rhizoma Heterosmilacis Japonicae) were separated and purified by column chromatography with silica gel, RP-C18, Sephadex LH-20, and preparative TLC. Their structures were elucidated on the basis of spectroscopic including IR, UV, MS(ESI-MS, HR-MS) and NMR(1H-NMR,13C-NMR, 1H-1H-COSY, HSQC, HMBC) and X-ray single crystal diffraction methods and chemical evidences.Results : 21 compounds have been isolated from water-soluble part of Rhizoma Heterosmilacis Japonicae. The structures of 18 compounds were elucidated based on their physicochemical and spectroscopic evidence as follow: Two Azoxyglycoside compounds : Macrozamin(1) and Cycasin(2); One amide compound: N-3-(3,5-dihydroxy-4,4-dimethyl-1-oxopentyl)-β-Alanine(3); Two Flavonoids: Hesperetin-7-0-rutimoside(4), Mangiferin(5); Three phenolic glycosides: 4-hydroxybenzoate glucoside(6) 3,5-dimethoxy-4-hydroxy-benzoic acid-O-β-D-glucoside(7), and glucosyringic acid(8); Nine phenolic compounds: 3,4,5-trihydroxy benzaldehyde(9), 3,4-dihydroxy benzaldehyde(10), 4-hydroxy benzoic acid(11), 3,4-dihydroxy benzoic acid(12), 2,5-dihydroxy-benzoic acid(13), 3,5-dimethoxyl-4-hydroxy benzoic acid(14), trans-Cinnamic acid(15), 4-ethyoxyl- Cinnamic acid(16), 4-(dimeth- oxymethyl)-phenol(17); One disaccharide: Sucrose.Conclusion:The systematic chemical constituents research was carried on the Rhizoma Heterosmilacis Japonicae. 18 compounds was isolated and indentified from the water-soluble part of it. It is wroth mentioning that all the compounds were firstly isolated from Rhizoma Heterosmilacis Japonicae except compound 1,4,5,6,7,8. In short, part of secondary metabolites of TCM Baituling had been figured out through the systematical chemical studies, which provide the chemical composition and theoretical basis for the control of quality standard and pharmacodynamic evaluation.