The chemical constituents of Helleborus thibetanus were isolated and purified by silica gel column chromatography, Sephadex LH-20 gel column chromatography, and semi-preparative RP-HPLC, and the structures of all compounds were identified by modern spectrographic technology(MS, NMR). The MTT method was used to measure the cytotoxicity of compounds 1-8. Twelve compounds were isolated from the roots and rhizomes of H. thibetanus and were identified as(25R)-22β,25-expoxy-26-[(O-β-D-glucopyranosyl)oxy]-1β,3β-dihydroxyfurosta-5-en(1), β-sitosterol myristate(2), β-sitosterol lactate(3), β-sitosterol 3-O-β-D-glucopyrannoside(4), 4,6,8-trihydroxy-3,4-dihydronaphthalen-1(2H)-one(5), 1,3,5-trimethoxybenzene(6), 7,8-dimethylbenzo pteridine-2,4(1H,3H)-dione(7), 1H-indole-3-carboxylic acid(8), p-hydroxy cinnamic acid(9), lauric acid(10), n-butyl α-L-arabinofuranoside(11) and methyl-α-D-fructofuranoside(12), respectively. Among them, compound 1 is a new compound and named thibetanoside L; compounds 2, 5-8, 11 are first isolated from the family Ranunculaceae; compound 12 is isolated from the genus Helleborus for the first time. The results of MTT assay showed that the IC_(50) values of compounds 1-8 against HepG2 and HCT116 cells were greater than 100 μmol·L~(-1).
Helleborus/chemistry* ; Molecular Structure ; Plant Roots/chemistry* ; Rhizome/chemistry* ; Magnetic Resonance Spectroscopy

OBJECTIVETo deal with the correlation between phylogeny, chemical constituents and pharmaceutical aspects of Ranunculaceae, namely a pharmaphylogenic study of this taxon.

METHODBased on chemical, pharmaceutical (both ethnopharmacologic and pharmacological) information, linking with different plant systems of Ranunculaceae.

RESULTChemical constituents of this taxon included several natural groups: benzylisoquinoline alkaloid, ranunculin, triterpenoid glycoside and diterpene alkaloid etc. Ranunculin and magoflorine were found to present simultaneously in some plants of this taxon.

CONCLUSIONCombining with therapeutic information, pharmaphylogenic research were in accordance with the phylogenetic system presented by Tamura that Ranunculaceae was proposed to be divided into six sub-families: Helleboroideae, Ranunculoideae, Cimicfugoideae, Isopyroideae, Thalictroideae and Coptidoideae. Results also supported the establishment of Cimicifugoideae.

Benzylisoquinolines ; isolation & purification ; Cimicifuga ; chemistry ; Diterpenes ; isolation & purification ; Furans ; isolation & purification ; Helleborus ; chemistry ; Methylglycosides ; isolation & purification ; Pharmacognosy ; classification ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; Ranunculaceae ; chemistry ; classification ; Triterpenes ; isolation & purification