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Bioactive oleanolic acid saponins and other constituents from the roots of Viguiera decurrens.

The bisdesmoside oleanolic acid saponin, 3-0-(methyl-beta-D-glucuronopyranosiduronoate)-28-0-beta-D-glucopyranosyl-oleanolate along with nine known compounds (two diterpenic acids, one chromene, three triterpenes, one steroidal glycoside, and two monodesmoside oleanolic acid saponins), were obtained from Viguiera decurrens roots. The chemical structure of the bisdesmoside oleanolic saponin was determined by chemical and NMR spectral evidence. A mixture of monodesmoside saponins displayed cytotoxic activity against P388 and COLON cell lines (ED50= 2.3 and 3.6 microg/ml, respectively). Two of the known compounds showed insecticidal activity against the Mexican bean beetle larvae (Epilachna varivestis).

Asteraceae↗

Cardioactive steroid saponins and other constituents from the aerial parts of Tribulus cistoides.

From the petrol extract of the aerial parts of Tribulus cistoides three steroid sapogenins and two N-acyltyramines were isolated, whereas the methanolic extract gave nine steroid saponins, among them the cardioactive cistocardin, saponin-3, saponin-4 and saponin-7. Furthermore, a furostanol diglycoside was isolated besides 5'-(hydroxysulphonyloxy) jasmonic acid, D-(+)-pinitol and sucrose. The structures were established by spectroscopic investigations of the isolated compounds and their hydrolysis products. 5'-(Hydroxysulphonyloxy) jasmonic acid has been prepared by partial synthesis.

Animals↗

Protective effect of iridals from saponin injury in Candida albicans cells.

Spirostanol saponin permeabilization of Candida albicans cells resulted in phosphate (Pi) leakage from these cells. Pre-incubation of C. albicans suspension cultures with ergosterol or cycloiridals inhibited Pi leakage when cells were subjected to saponins. Saponins were shown to precipitate ergosterol but not cycloiridals. Inhibition of Pi leakage in the presence of cycloiridals could thus be due to their incorporation into cell membranes. Cycloiridals, which were previously shown to modify fluidity parameters in artificial phospholipid bilayers, seem in vivo to have a protective effect on membranes against surfactant (saponin) injury.

Biological Transport↗

Triterpenoid saponins from Berneuxia thebetica.

Four triterpenoid saponins were isolated from Berneuxia thibetica. On the basis of chemical and spectroscopic evidence, three new saponins, berneuxia saponins A, B and C, were elucidated as 21-tigloylbarringtogenol C-3 beta-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-galactopyranosyl(1-->3)[bet a- D-glucopyranosyl(1-->2)-beta-D-glucuronopyranoside], 28-tigloylbarringtogenol C-3 beta-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-galactopyranosyl(1-->3)[bet a- D-glucopyranosyl(1-->2)-beta-D-glucuronopyranoside] and 16 alpha-28-dihydroxyolean-12-en-21-one-3-O-alpha-L-rhamnopyranosyl(1 -->2) -beta-D-galactopyranosyl(1-->3)[beta-D-glucopyranosyl(1-->2)-beta-D- glucuronopyranoside]. The known saponin was desacyl jegosaponin.

Carbohydrate Conformation↗

Steroidal saponins from the bulbs of Lilium candidum.

Five new spirostanol saponins and a new furostanol saponin were isolated from the fresh bulbs of Lilium candidum. Their structures were elucidated on the basis of spectroscopic analysis, including two-dimensional NMR spectroscopic techniques and the result of acid hydrolysis. The isolated saponins contained a branched triglycoside moiety assigned as O-alpha-L-rhamnopyranosyl-(1-->2)-O-[beta-D-glucopyranosyl-(1-->6)]-beta - D-glucopyranose with the formation of an O-glycosidic linkage to C-3 of the aglycone as the common structural feature. The inhibitory activity of the saponins on Na+/K+ ATPase was evaluated.

Carbohydrate Conformation↗

Suppressive effects of vietnamese ginseng saponin and its major component majonoside-R2 on psychological stress-induced enhancement of lipid peroxidation in the mouse brain.

We investigated the in vivo effects of Vietnamese ginseng saponin (VG saponin) and its major component majonoside-R2 (MR2) on psychological stress-induced enhancement of lipid peroxidation in the mouse brain. Psychological stress exposure using a communication box system for 4 h significantly increased the content of thiobarbituric acid reactive substance (TBARS), an index of lipid peroxidation activity, in the brain. Pretreatment with VG saponin (15-25 mg/kg, PO) and MR2 (1-10 mg/kg, IP) significantly attenuated the psychological stress-induced increase in TBARS content in the brain. The aglycone of MR2 (MR2-aglycone: 1.2 mg/kg, IP), at the equivalent dose of MR2 (i.e., 3 mg/kg, IP), also produced the suppressive effect on the increase in the TBARS content. The in vivo suppressive effect of MR2 was dose dependently attenuated by flumazenil (3 and 10 mg/kg, IP), a benzodiazepine receptor antagonist, and pregnenolone sulfate (10 mg/kg, IP), a neurosteroidal negative allosteric modulator of GABA(A) receptors. These findings suggest that VG saponin and its major component MR2 have preventive effects on the psychological stress-induced brain cell membrane damage, and that the effect of MR2 is partly due to enhancement of GABA(A)-ergic systems in the brain.

Animals↗

Adjuvant effects of orally administered saponins on humoral and cellular immune responses in mice.

We have described adjuvant effects of orally administered Quillaja saponins on the immune responses of mice fed inactivated rabies antigen (AG). The in vivo lymphocyte proliferation in mice fed antigen + saponin (AG + SAP) was significantly greater than that in mice fed antigen (AG) alone. Further, the mitogen-induced cell proliferative responses in animals primed with AG + SAP was markedly increased compared with those in the AG group. These changes in clonal expansion were associated with an enhanced helper T cell (Th) and B cell co-operation. The in vivo cell proliferation and in vitro mitogen-induced responses of mice fed AG + SAP correlated with enhanced antibody synthesis. In mice fed saponin alone, there were significant increases in clonal expansion and lymphocyte function. Our present data indicate that the immunocompetence in animals fed AG + SAP was indeed evoked by saponins. Cytotoxic T lymphocyte activity in mice fed SAP or AG + SAP was detected 7 days after booster, in contrast to 21 days in mice fed AG alone. The natural killer cell activity in mice fed SAP alone was greatly enhanced and persisted for an extended period of time.

Adjuvants, Immunologic↗

Saponins, IL12 and BCG adjuvant in the FML-vaccine formulation against murine visceral leishmaniasis.

The FML antigen of Leishmania donovani, in combination with either Riedel de Haën (R), QuilA, QS21 saponins, IL12 or BCG, was used in vaccination of an outbred murine model against visceral leishmaniasis (VL). Significant and specific increases in anti-FML IgG and IgM responses were detected for all adjuvants, and in anti-FML IgG1, IgG2a and IgG2b and delayed type of hypersensitivity to L. donovani lysate (DTH), only for all saponins and IL12. The QS21-FML and QuilA-FML groups achieved the highest IgG2a response. QuilA-FML developed the strongest DTH and QS21-FML animals showed the highest serum IFN-gamma concentrations. The reduction of parasitic load in the liver in response to each FML-vaccine formulation was: 52% (P<0.025) for BCG-FML, 73% (P<0.005) for R-FML, 93% (P<0.005) for QuilA-FML and 79.2% (P<0.025) for QS21-FML treated animals, respectively. Protection was specific for R-FML and QS21-FML while the QuilA saponin treatment itself induced 69% of LDU reduction. The FML-saponin vaccines promote significant, specific and strong protective effects against murine visceral leishmaniasis. BCG-FML induced minor and non-specific protection while IL12-FML, although enhancing the specific antibody and IDR response, failed to reduce the parasitic load of infected animals.

Adjuvants, Immunologic↗

The use of commercial saponin from Quillaja saponaria bark as a natural larvicidal agent against Aedes aegypti and Culex pipiens.

The larvicidal activity of commercial bark saponin extract (Sigma) from Quillaja saponaria was studied on 3rd-4th instar larvae of Aedes aegypti and Culex pipiens (vectors for dengue fever and Western Nile virus, respectively). The larvae were exposed to serial concentrations (1000, 800, 500, 300, 100, 10, 1, 0.1 and 0.01 mg/l) of the extract for 1, 3, 5, 7 and 11 days. The results indicate that commercial bark saponin is toxic, causing 100% larval mortality in A. aegypti and C. pipiens after 1 and 5 days at a dosage of 800 and 1000 mg/l, respectively. Interestingly, while bark saponins had a toxic effect on larvae there was no effect on egg hatchability in either species. The results obtained suggest that, in addition to their known activities, saponins can also serve as natural larvicidal compounds.

Aedes↗

Protective effects of steroid saponins from Paris polyphylla var. yunnanensis on ethanol- or indomethacin-induced gastric mucosal lesions in rats: structural requirement for activity and mode of action.

The methanolic extract from the rhizomes of Paris polyphylla SM. var. yunnanensis (FR.) H-M. was found to potently inhibit ethanol-induced gastric lesions in rats. Through bioassay-guided separation, four known spirostanol-type steroid saponins, pennogenin 3-O-alpha-L-rhamnopyranosyl(1-->2)-[alpha-L-arabinofuranosyl(1-->4)]-beta-D-glucopyranoside (1), pennogenin 3-O-alpha-L-rhamnopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->4)-[alpha-L-rhamnopyranosyl(1-->2)]-beta-D-glucopyranoside (2), diosgenin 3-O-alpha-L-rhamnopyranosyl(1-->2)-[alpha-L-arabinofuranosyl(1-->4)]-beta-D-glucopyranoside (3), and diosgenin 3-O-alpha-L-rhamnopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->4)-[alpha-L-rhamnopyranosyl(1-->2)]-beta-D-glucopyranoside (4), and a new furostanol-type steroid saponin, parisaponin I (5), together with two known furostanol-type steroid saponins, trigofoenoside A (6) and protogracillin (7), were isolated from the active fraction. Compounds 1-4 (1.25-10 mg/kg, po) strongly inhibited gastric lesions induced by ethanol and indomethacin. With regard to structural requirement of steroid saponins, the 3-O-glycoside moiety and spirostanol structure were found to be essential for the activity and the 17-hydroxyl group in the aglycon part enhanced the protective effects against ethanol-induced gastric lesions. The protective effects of 1 and 3 against ethanol-induced gastric lesions were attenuated by pretreatment with indomethacin and N-ethylmaleimide. Compounds 1 and 3 weakly inhibited acid secretions in pylorus-ligated rats. These findings suggested that endogenous prostaglandins and sulfhydryl compounds were involved in the protective activity.

Animals↗

Solid-phase extraction NMR studies of chromatographic fractions of saponins from Quillaja saponaria.

The saponin mixture QH-B from the tree Quillaja saponaria var. Molina was fractionated by RP-HPLC in several steps. The fractions were analyzed by solid-phase extraction NMR (SPE-NMR), a technique combining the workup by solid-phase extraction with on-line coupling to an NMR flow probe. Together with MALDI-TOF mass spectrometry and comparison with chemical shifts of similar saponins, the structures of both major and minor components in QH-B could be obtained. The procedure described is a simple method to determine the structure of components in a complex mixture. The two major fractions of the mixture were found to contain at least 28 saponins, differing in the carbohydrate substructures. Eight of these have not previously been determined. The 28 saponins formed 14 equilibrium pairs by the migration of an O-acyl group between two adjacent positions on a fucosyl residue.

Adjuvants, Immunologic↗

Improvement of Panax notoginseng cell culture for production of ginseng saponin and polysaccharide by high density cultivation in pneumatically agitated bioreactors.

A Panax notoginseng cell culture was successfully scaled up from shake flask to 1.0-L bubble column reactor and concentric-tube airlift reactor. High-density bioreactor batch cultivation was carried out using a modified MS medium. The maximum cell density in batch cultures reached 20.1, 21.0 and 24.1 g/L in the shake flask, bubble column and airlift reactors, respectively, and their corresponding biomass productivity was 950, 1140 and 1350 mg/(L x d) for each. The productivity of ginseng saponin was 70, 96 and 99 mg/(L x d) in the flask, bubble column and airlift reactors, respectively; and the polysaccharide productivity reached 104, 119 and 151 mg/(L x d) for each. Furthermore, a fed-batch cultivation strategy was developed on the basis of specific oxygen uptake rate (SOUR), i.e., sucrose feeding before a sharp decrease of SOUR, and the highest cell density of 29.7 g/L was successfully achieved in the airlift bioreactor on day 17 with a very high biomass productivity of 1520 mg/(L x d). The concentrations of ginseng saponin and polysaccharide reached about 2.1 and 3.0 g/L, respectively, and their productivity was 106 (saponin) and 158 mg/(L x d) (polysaccharide). This work successfully demonstrated the high-density bioreactor cultivation of P. notoginseng cells in pneumatically agitated bioreactors and the reproduction of the shake flask culture results in bioreactors. The cell density, biomass productivity, production titer and productivity of both ginseng saponin and polysaccharide obtained here were the highest that have been reported on a reactor scale for all the ginseng species.

Biomass↗

Triterpene saponins from aerial parts of Medicago arabica L.

Eight major triterpene saponins have been isolated from the aerial parts of Medicago arabica and their structures elucidated by FAB-MS and NMR analysis. Three of them are new compounds and are identified as 3-O-(alpha-L-arabinopyranoside) bayogenin, 3-O-(alpha-L-arabinopyranosyl), 28-O-(beta-D-glucopyranoside) bayogenin, and 3-O-[alpha-L-arabinopyranosyl(1-->2)-beta-D-glucuronopyranosyl], 28-O-beta-D-glucopyranoside 2-beta-hydroxyoleanolic acid. Two saponins, identified as 3-O-(alpha-L-arabinopyranoside) hederagenin and 3-O-(alpha-L-arabinopyranosyl), 28-O-(beta-D-glucopyranoside) hederagenin are known compounds but not previously reported as saponin constituents of Medicago species, while three other saponins, being mono- and bidesmosides of hederagenin, have been previously isolated from roots of M. sativa.

Hydrolysis↗

Triterpene saponins from the roots of Medicago hybrida.

Fourteen triterpene saponins (1-14) have been isolated from the roots of Medicago hybrida and their structures elucidated by FAB-MS and NMR analysis. Two of them are new compounds and were identified as hederagenin 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (7) and oleanolic acid 3-O-[beta-D-galactopyranosyl(1-->2)-beta-D-glucuronopyranosyl]-28-O-[alpha-L-rhamnopyranosyl(1-->4)-beta-D-glucopyranoside] (14). Seven saponins being mono- and bidesmosides of hederagenin (1, 5, 6, 9), one bidesmoside of bayogenin (2), and two bidesmosides of 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid (11) and oleanolic acid (13) are known compounds but not previously reported as saponin constituents of Medicago, whereas five other saponins, being mono- and bidesmosides of medicagenic acid (3, 4, 8, 10, 12), and one monodesmoside of hederagenin (8) have been previously isolated from other Medicago species. The presence of 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid might represent an interesting intermediate in the biosynthesis of these substances.

Magnetic Resonance Spectroscopy↗

Facile synthesis of saponins containing 2,3-branched oligosaccharides by using partially protected glycosyl donors.

Two natural saponins 1 and 2, isolated from Solanum indicum L., and containing 2,3-branched sugar moieties, have been efficiently synthesized. Partially protected monosaccharide and disaccharide donors were used to facilitate target synthesis. Stereo factors were critical in incorporating 2,3-branched sugars on steroid aglycones. Saponin 1 was synthesized in five steps and 30% overall yield, while saponin 2 was obtained using six straightforward sequential reactions in 31% overall yield. Saponin 2 shows promising cytotoxic activity toward human hepatocellular carcinoma BEL-7402 with an IC50 of <6 microg/mL.

Antineoplastic Agents, Phytogenic↗

Steroidal saponins from Dracaena surculosa.

A phytochemical investigation of the whole plant of Dracaena surculosa resulted in the isolation of nine steroidal saponins, including three new bisdesmosidic spirostanol saponins, named surculosides A (1), B (2), and C (3), and a new bisdesmosidic furostanol saponin (4), which are based on (25S)-spirost-5-ene-1beta, 3beta-diol [(25S)-ruscogenin] as the aglycon. The structures of 1-4 were determined by spectroscopic analysis, including 2D NMR spectroscopic data, and the results of hydrolytic cleavage. The isolated saponins were evaluated for their cytotoxic activity against HL-60 human promyelocytic leukemia cells.

Carbohydrate Conformation↗

Triterpenoid saponins from the fruits of Caryocar villosum.

Fourteen new triterpenoid saponins (1-14) were isolated from the methanol extract of the fruits of Caryocar villosum along with 10 known saponins. Their structures were established on the basis of extensive NMR (1H, 13C, COSY, TOCSY, ROESY, HSQC, and HMBC) and ESIMS studies. The toxicity of the methanolic extracts of the peel and the pulp of fruits and the crude saponin fraction of the peel was assessed using the Artemia salina test. The antimicrobial activities of caryocarosides IV-21 (14), II-1 (16), III-1 (17), and IV-9 (20) and of saponin 23 were also studied in vitro on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Mycobacterium smegmatis, and Enterococcus faecalis bacteria.

Animals↗

Triterpene saponins from clematis mandshurica.

Four new triterpene saponins, clematomandshurica saponins A-D (1-4), together with three known saponins (5-7) have been isolated from the roots and rhizomes of Clematis mandshurica. Their structures were elucidated on the basis of their spectroscopic evidence and hydrolysis. Clematomandshurica saponins A and B showed significant inhibitory activity on cyclooxygenase-2 (IC50 = 2.66 and 2.58 microM, respectively).

Animals↗