Studies On The Saponin Constituents Of Anemone Tomentosa

Anemone tomentosa, a perennial herbaceous plants, belongs toRanunculaceae. The plants was investigated, a widely occurring perennial herb inShaanxi, Henan, Hebei, Gansu and Sichuan provinces of China. Its root has beenused in traditional Chinese medicines for the treatment of dysentery, malaria,infantile malnutrition, carbuncles, etc. Pharmacological research indicates thatplants of this genus have many kinds of activities, including antipyretic andanalgesia, antibacterial and anti-inflammation, antitumor activity. Saponins arethe main bioactive compounds.In this paper, the ethanol extract of the roots of Anemone tomentosa waschromatographed on the silica gel column, D-101macroporous adsorption resin,reversed-phase ODS, Sephadex LH-20and SP-HPLC to yield15compounds.The structures of13compounds were identified through ESI-MS,1H-NMR,13C-NMR, DEPT, HSQC, HMBC, NOESY, TOCSY,1H-1H COSY technologyand chemical evidence, including4new compounds and9known compounds.Which are respectively:7-ethyl-coumarin (1),3-O-α-L-arabinopyranosyl-(1→2)-β-D-xylopyranosyloleanolic acid28-O-β-D-glucopyransyl-(1→4)-β-D-glucopy- ranoside (2), koreanaside1(3), hederasaponin B (4), clematiganoside A (5),huzhangoside D (6), huzhangoside D (7),3-O-β-D-galactopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-β-D-xylopyranosyl oleanane acid28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside(8),3-O-β-D-ribopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-β-D-xylopyranosyl hederagenin28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside(9),3-O-β-D-ribopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→4)]-α-L-arabinopyranosyl hederagenin28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (10), clematichinenoside A (11), β-sitosterol (12),β-daucosterol (13). Compounds2,8,9and10are new compounds andcompounds3,5and11are isolated for the first time from this genus. Four newcompounds are identified to be oleanane type triterpenoid saponins. Compound8that have D-galactopyranosyl group was found for the second time from thisgenus. Got5second glycosides through alkaline hydrolysis. Which arerespectively: CP2(4a), clemargenoside G (5a), clemontanoside C (6a), CP4(7a)and3-O-β-D-ribopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-[β-D-glucopy-ranosyl-(1→4)]-α-L-arabinopyranosyl hederagenin(10a). Compound10a is anew compound that was named as tomentoside Aa.Ten triterpenoid saponins (2-11) isolated from the roots of anemonetomentosa have been tested on anticancer activities against three different tumorcell lines. The result showed that oleanane saponins of two suger groupexhibited inactive cytotoxicity against all the three cancer cell lines. But5second glycosides (4a-7a,10a) through alkaline hydrolysis have forceful activecytotoxicity which of7a has the most forceul. This thesis elucidated thestructure-activity relationship of oleanane saponion and got a clear conclusion. Innovations and significance:1. Took an in-depth study of the chemical constituents from Anemonetomentosa, identified the structures of13compounds and got5secondglycosides through chemical methods. These work enlarged our knowledgesabout the derivatives of Anemone tomentosa.2. We found five new compounds including four native glycosides and onesecond glycoside. Saponin8of D-galactopyranosyl group was found for thesecond time from this genus. As to saponins2and3which were composed bytwo pepentaglucoses group were the first find in this genus.3. We found5anti-cancer compounds by systemically tumor cells toxocityscreening tests, and discussed structure-efficiency relationship throughexperiments. All these data would provide guidances for developing this type ofsaponin.4. We got5derivatives through alkaline hydolysis method which suggestedthat we can get actived compound through this similar chemical method. Thismethod will offer a new direciton for developing activating compounds, and it’sa new attempt to developing activating saponins.5. Anemone tomentosa distribute widely in China. The present study offeredscientific basis for utilizing this abundant plant resource. We have systemeticallystudied Anemone taipaiensis and Anemone rivularis var. flore-minore. Thepresent study offered new information for furthering study of this genus anddeepening our knowledge about this genus. Also, this study provided reasonableground for its further exploitation this species of Anemone, and offered valuableleads for pursuing new antitumor drugs.