Gland Stem Swine Siegesbeckiae Antagonistic Proinflammatory Factor Active Ingredients

Siegesbeckia pubescens Makino belongs to the genus Siegesbeckia and the family Compositae. They have been used as a traditional Chinese medicine to treat rheumatic arthritis, hypertension, malaria, etc. It is reported in Dictionary of Chinese Material Medica and Ch P (2005) in China. In order to elucidate the active principle of S. pubescens, we have investigated it on the NO production and TNF-αrelease induced by LPS. Through bioassay-guided fractionation, fifteen compounds were isolated from S. pubescens, by means of chormatography method, eleven compounds are identified by application of special evidence, such as chemical methods and comparison with some authentic samples, standard spectra, physical-chemical data, and these compounds are established as follows: tetracosa-carbonic acid (1),β-sitosterol (2), ursolic acid (3), N-(N-benzoyl-L-phenylalanyl)-O-actyl-L-phenylalanol (4), bis(2-ethylhexyl)phthalate (5), 6-Hydroxy-2-naphthoic acid (6), stigmasteryl-3-O-β-D-glucopyranoside (7), ent-16β,17-dihydroxy- kauran-19-oic acid (8), darutigenol (9), uracil (10), kirenol (11). For the first time, Compounds (3)-(6) and (10) are obtained from Siegesbeckia, and compound (7) is firstly separated from Siegesbeckia pubescens.In the course of structural determination, extensive 2D-NMR techniques, especially HMBC (1H-detected multiple-bond heteronuclear multiple quantum coherence) experiments, are successfully applied to determine the structure of compounds. 1H and 13C-signals of compounds are assigned firstly by means of 2D-NMR spectra. Some mistakes which occurred in the literatures of compounds are properly corrected.With respect to the bioassay, we indicate the compounds (1)-(3), (5), (9) were found to have strong inhibitory effects on the NO production, IC50 values are 13.1μg/mL, 13.9μg/mL, 5.9μg/mL, 10.0μg/mL, 6.2μg/mL, respectively. And all the IC50 values were better than the positive control of Hydrocortisone. (3) is the most active. Compounds (3)-(10) showed strong inhibitory activities on TNF-αrelease, IC50 are 1.6 μg/mL, 2.9μg/mL, 2.9μg/mL, 4.3μg/mL, 4.2μg/mL, 4.4μg/mL, 0.8μg/mL, 3.6μg/mL. 9 showed the IC50 values lower than 1μg/mL.