Investigation On Anti-Influenza Virus Activities Of Five Formulas Of Zhuangyao Medicine And Chemical Constituents From The Seeds Of Caesalpinia Minax Hance

Five formulas of ZhuangYao medicine(including formula 1:Helictercs angustifolia L.,Artemisia annua L.,etc,2:Clausena lansium(Lour.)Skeels., Morus alba L.,etc,3:Euphoria longan L.,Urena lobata L.,etc,4:Gaultheria yunnanensis(Franch.)Rehd.5:ku-shi-lian)were extracted with ethanol.The anti-influenza virus activities of the crude extracta siccas were tested in vitro by cytopathic effect assay(CPE).The formula 1,2 and 5 showed anti-influenza activities against virus A(H3N2)with the IC₅₀values of 577.35μg/ml,429.5μg/ml and 429.5μg/ml.According to the results,the different polarity fractions of the formula 2 and 5(such as petroleum ether fraction,chloroform fraction, ethyl acetate fraction,etc.)were further investigated,the chloroform fractions of formula 2 and formula 5,the petroleum ether fraction of formula 5 were confirmed to be effective with the IC₅₀values of 333.3μg/ml,333.3μg/ml and 111.1μg/ml.Caesalpinia minax Hance is a plant belonging to the genus Caesalpinia L.. The seeds of it(ku-shi-lian in Chinese)have been used as a folk remedy for the treatment of common cold,dysentery,rheumatism,influenza and hepatitis.In this paper,chloroform and ethyl acetate fractions from the ethanol extract were researched for searching the anti-influenza compounds.Nine compounds were isolated from ku-shi-lian by various methods including repeated silica gel column chromatography,sephadex LH-20 column chromatography and preparative HPLC.By means of spectral data,comparisons with authentic compounds and known data from literatures,the structures of seven compounds were identified asβ-Sitosterol(â… ),5-hydroxymethyl-2-furfuraldehyde(â…¡),Neocaesalpin B(â…¢), Neocaesalpin L(â…£),Neocaesalpin MO(â…¤),Neocaesalpin MP(â…¥),Daucosterol (â…¦).Among them,compoundsâ…¤andâ…¥were new cassane diterpenoids lactone;compoundâ…¡was obtained from the genus Caesalpinia L.for the first time;compoundsâ… andâ…¢were isolated from C.minax.for the first time...