Saponins from Chinese folk medicine, "zhu jie xiang fu," Anemone raddeana REGEL

From the Chinese folk medicine "Zhu jie xian fu" (roots of Anemone raddeana REGEL, Ranunculaceae), two new oleanane-type glycosides, named raddeanosides R8 (1) and R9 (2), were isolated. The structures of 1 and 2 were determined as 3-O-alpha-L-rhamnopyranosyl-(1----2)-O-beta-D-glucopyranosyl- (1----2)-alpha-L-arabinopyranosyl oleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1----4)-O-beta-D-glucopyranosyl-(1----6)-b eta-D- glucopyranoside and 3-O-alpha-L-rhamnopyranosyl-(1----2)-O-beta-D-glucopyranosyl-(1----2)-al pha-L- arabinopyranosyl 27-hydroxyoleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1----4)-O-beta-D-glucopyranosyl-(1----6)-b eta-D- glucopyranoside, respectively.     

Studies on the constituents of Luffa acutangula Roxb. I. Structures of acutosides A--G, oleanane-type triterpene saponins isolated from the herb.

From the herb of Luffa acutangula ROXB. (Cucurbitaceae), seven oleanane-type triterpene saponins, acutosides A--G, were isolated and their structures were determined. Acutoside A is oleanolic acid 3-O-beta-D-glucopyranosyl-(1----2)-beta-D-glucopyranoside. Acutosides B, D, E, F and G have a common prosapogenin structure, acutoside A, and only differ in the structures of the ester-linked sugar moieties. Acutoside B is a 28-O-[O-beta-D-xylopyranosyl-(1----4)-O-alpha-L-rhamnopyranosyl-(1----2) -alpha-L-arabinopyranosyl] ester, D is a 28-O-[O-beta-D-xylopyranosyl-(1----3)-O-beta-D-xylopyranosyl-(1----4)-O- alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester, E is a 28-O-[O-alpha-L-arabinopyranosyl-(1----3)-O-beta-D-xylopyranosyl-( 1----4)-O-alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester, F is a 28-O-[O-beta-D-xylopyranosyl-(1----3)-[O-beta-D-xylopyranosyl-(1----4)-O -alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester, and G is a 28-O-beta-D-xylopyranosyl-(1----3)-[O-alpha-L-arabinopyranosyl-(1- ---3)-O-beta-D-xylopyranosyl-(1----4)]-O-alpha-L- rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester. Acutoside C is a machaelinic acid (=21 beta-hydroxyoleanolic acid) saponin having the same sugar moiety as that of acutoside B.     

Steroidal saponins and alkaloids from the bulbs of Lilium brownii var. colchesteri

The fresh bulbs of Lilium brownii var. colchesteri were found to contain five steroidal saponins: 26-O-beta-D-glucopyranosylnuatigenin 3-O-alpha-L-rhamnopyranosyl-(1----2)-beta-D-glucopyranoside (6), 26-O-beta-D-glucopyranosylnuatigenin 3-O-alpha-L-rhamnopyranosyl-(1----2)-O-[beta-D-glucopyranosyl-(1----4)]- beta-D-glucopyranoside (7), brownioside (8), deacylbrownioside (9) and 27-O-(3-hydroxy-3-methylglutaroyl)isonarthogenin 3-O-alpha-L-rhamnopyranosyl-(1----2)-O-[beta-D-glucopyranosyl-(1----4)- beta-D-glucopyranoside (10); and two steroidal alkaloids: beta 1-solamargine (11) and solasodine 3-O-alpha-L-rhamnopyranosyl-(1----2)-O-[beta-D-glucopyranosyl-(1----4)- beta-D-glucopyranoside (12); along with several phenolic constituents. Compounds 7, 10 and 12 are new naturally-occurring compounds.     

High-performance liquid chromatographic identification of disaccharides generated from heparan sulphate isomers using heparitinases

Specific heparan sulphate-lyases, heparitinases I and II, were used to identify unsaturated disaccharide constituents generated from heterogeneous heparan sulphate isomers. All determinations were made using high-performance liquid chromatography with a column containing a sulphonized styrene-divinylbenzene copolymer. Unsaturated disaccharides generated from variously sulphated heparan sulphate isomers after simultaneous digestion with heparitinases I and II facilitated separation of the individual disaccharides, based on sulphate groups at the specific position of the uronic acid and glucosamine residues. The simultaneous digestion with heparitinases I and II produces unsaturated disaccharides from heparan sulphate isomers with the structure of 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-amino-deoxy-D-glucose, 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-deoxy-2-sulphamido-D-glucose, 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-aminodeoxy-6-O-sulpho-D-glucose, 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-deoxy-2-sulphamido-6-O-sulpho-D-glucose, 4-deoxy-2-O-sulpho-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-amino-2-deoxy-6-O-sulpho-D-glucose and 4-deoxy-2-O-sulpho-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-deoxy-2-sulphamido-6-O-sulpho-D-glucose.     

Studies on the constituents of Aster scaber Thunb. III. Structures of scaberosides B7, B8 and B9, minor oleanolic acid glycosides isolated from the root.

Three new oleanolic acid 3,28-O-bisdesmosides, scaberosides B7, B8 and B9, were isolated as minor saponins from the root of Aster scaber THUNB. (Compositae), and their structures were determined based on spectral and chemical evidence as follows. Scaberoside B7 is 3-O-beta-D-glucopyranosyluronic acid oleanolic acid 28-[O-beta-D-apiofuranosyl-(1----3)-[O-beta-D-xylopyranosyl-(1---- 4)-O-alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranosyl] ester, scaberoside B8, 3-O-beta-D-glucopyranosyl oleanolic acid 28-[O-beta-D-xylopyranosyl-(1----4)-O-alpha-L-rhamnopyranosyl-(1----2)-a lpha-L-arabinopyranosyl] ester, and scaberoside B9, 3-O-beta-D-glucopyranosyluronic acid oleanolic acid 28-[O-alpha-L-rhamnopyranosyl-(1----2)-[O-beta-D-xylopyranosyl-(1----6)] -beta-D-glucopyranosyl] ester. Scaberosides B7 and B9 were obtained as their methyl esters.     

Structures of two new fibrinolytic saponins from the seed of Luffa cylindrica Roem

Two new fibrinolytic saponins, lucyosides N and P, were isolated from the seeds of Luffa cylindrica Roem. (Cucurbitaceae). On the basis of chemical and spectral evidence, lucyoside N was characterized as 3-O-beta-D-galactopyranosyl-(1----2)-beta-D-glucuronopyranosyl-28- O-beta-D-xylopyranosyl-(1----4)-[beta-D-glucopyranosyl-(1----3)]-alpha-L -rhamnopyranosyl-(1----2)-alpha-arabinopyranosyl quillaic acid. Lucyoside P was characterized as a gypsogenin glycoside with the same sugar moiety as lucyoside N.     

Periandradulcins A, B and C: phosphodiesterase inhibitors from Periandra dulcis Mart

During the course of our screening of bioactive natural products, three new saponins named periandradulcins A (1), B (2) and C (3) were isolated as phosphodiesterase (PDE, EC 3.1.4.17) inhibitors from 80% MeOH extract of the roots of Periandra dulcis Mart. (Leguminosae) by a combination of column chromatography and reversed- and normal-phase high-performance liquid chromatography (HPLC). On the basis of 1H-, 13C- and two-dimensional nuclear magnetic resonance (NMR) spectral data and chemical evidence, their chemical structures were characterized as 3-O-beta-[alpha-L-rhamnopyranosyl(1----2)-beta-D-xylopyranosyl(1----2)-b eta-D- glucuronopyranosyl]-30-hydroxyl-25-formylolean-18-ene-22 beta-O-syringate, 3-O-beta-[alpha-L-rhamnopyranosyl(1----2)-beta-D- xylopyranosyl(1----2)-beta-D-glucuronopyranosyl]-22 beta-hydroxyl-25- formylolean-12-ene and 3-O-beta-[alpha-L-rhamnopyranosyl(1----2)-beta-D- glucopyranosyl(1----2)-beta-D-glucuronopyranosyl]-22 beta-hydroxyl-25-formylolean-18-ene, respectively. The concentrations of periandradulcins A, B and C required to give 50% inhibition (IC50 values) of PDE from bovine heart, were 0.033, 7.6 and 7.7 microM, respectively. Compound 1 was the most potent among the known PDE inhibitors; it inhibited PDE-I (IC50:0.0022 microM) twenty and forty times more effectively than PDE-II and -III, respectively.     

Structures of two new steroidal glycosides, soladulcosides A and B from Solanum dulcamara

The structures of two new steroidal glycosides named soladulcosides A and B, isolated from the aerial parts of Solanum dulcamara including new sapogenols, were elucidated as (22R, 25R)-3 beta, 15 alpha, 23 alpha-trihydroxy-5 alpha-spirostan-26-one 3-O-alpha-L-rhamnopyranosyl-(1----2)-beta-D-glucopyranoside and (22R, 25R)-3 beta,23 alpha-dihydroxy-5 alpha-spirostan-26-one 3-O-alpha-L-rhamnopyranosyl-(1----2)-[alpha-L-rhamnopyranosyl-(1----4)]- beta-D-glucopyranoside, respectively.     

Indonesian medicinal plants. I. Chemical structures of calotroposides A and B, two new oxypregnane-oligoglycosides from the root of Calotropis gigantea (Asclepiadaceae).

Two new oxypregnane-oligoglycosides named calotroposides A (1) and B (2) have been isolated from the root of Calotropis gigantea (Asclepiadaceae), an Indonesian medicinal plant, and their chemical structures have been elucidated by chemical and spectroscopic methods as 12-O-benzoyllineolon 3-O-beta-D-cymaropyranosyl(1----4)-beta-D-oleandropyranosyl( 1----4)- beta-D-oleandropyranosyl(1----4)-beta-D-cymaropyranosyl(1--- -4)-beta-D- cymaropyranoside and 12-O-benzoyldeacetylmetaplexigenin 3-O-beta-D-cymaropyranosyl(1---4)-beta-D-oleandropyranosyl(- ---4)- beta-D-oleandropyranosyl(1----4)-beta-D-cymaropyranosyl(1--- -4)- beta-D-cymaropyranoside, respectively.     

Saponins from Amaranthus hypochondriacus.

Four triterpenoid saponins were isolated from Amaranthus hypochondriacus which are grain crops in the Nepal, Mexico and South America. Their structures were elucidated based on spectral evidence to be: (1) 3-O-alpha-L-rhamnopyranosyl(1----3)-beta-D-glucuronopyranosyl-2 beta,3 beta-dihydroxyolean-12-en-28-oic acid 28-O-beta-D-glucopyranosyl ester. (2) 3-O-alpha-L-rhamnopyranosyl(1----3)-beta-D-glucuronopyranosyl-2 beta,3 beta- dihydroxyolean-12-en-23-al-28-oic acid 28-O-beta-D-glucopyranosyl ester; (3) 3-O-alpha-L-rhamnopyranosyl(1----3)-beta-D-glucuronopyranosyl-2 beta, 3 beta-dihydroxy-30-norolean-12,20(29)-dien-28-oic acid 28-O-beta-D-glucopyranosyl ester. (4) 3-O-alpha-L-rhamnopyranosyl (1----3)-beta-D-glucuronopyranosyl-2 beta, 3 beta-dihydroxy-30-norolean-12, 20(29)-dien-23-al-28-oic acid 28-O-beta-D-glucopyranosyl ester.     

Discrimination between endotoxin and (1----3)-beta-D-glucan using turbidimetric kinetic assay with Limulus amebocyte lysate

A procedure for the Limulus amebocyte lysate (LAL) test discriminating between endotoxin and (1----3)-beta-D-glucan based on the turbidimetric kinetic method was proposed. Endotoxin and (1----3)-beta-D-glucan, which are elicitors of the activation of LAL, showed different reaction courses with this lysate. To analyze the difference in the reactions, two parameters, the maximum differential coefficient of the reaction (Dmax) and the reaction time required to obtain Dmax (Tp) were defined. The logarithmic plottings of Tp versus Dmax (Tp-Dmax plot) discriminated between endotoxin and (1----3)-beta-D-glucan. Endotoxin was measured with a standard curve plotting logarithmic endotoxin concentration versus Dmax (ET-Dmax plot). The endotoxin calculated from Dmax was less influenced by (1----3)-beta-D-glucan than that calculated from the usual gelation time. A small amount of endotoxin in a sample could be concealed by the addition of polymyxin B, which inhibited the activation of LAL by endotoxin. (1----3)-beta-D-glucan was measured without being affected by the presence of a small amount of endotoxin using LAL with polymyxin B. The following procedure is proposed as a LAL test to discriminate between endotoxin and (1----3)-beta-D-glucan. (1) Identify the main substance (endotoxin or (1----3)-beta-D-glucan) triggering the activation of LAL using the Tp-Dmax plot. (2) Use the appropriate method to measure the main substance: the ET-Dmax plot for endotoxin or the LAL with polymyxin B for (1----3)-beta-D-glucan.     

High-performance anion-exchange chromatography of homogeneous D-gluco-oligosaccharides and -polysaccharides (polymerization degree greater than or equal to 50) with pulsed amperometric detection

High-performance anion-exchange chromatography under alkaline conditions with pulsed amperometric detection was applied to the analyses of (1----2)-, (1----3)-, (1----4)- and (1----6)-linked homogeneous alpha- or beta-D-gluco-oligosaccharides and -polysaccharides up to a degree of polymerization (DP) of greater than or equal to 50. Each series of homogeneous D-gluco-oligomers and -polymers showed a linear relationship between log k' and DP in isocratic elution using 150 mM sodium hydroxide solution containing 100 mM sodium acetate as the eluent. An effective separation of individual members of an homologous series of linear glucans was achieved using gradient elution, accomplished by maintaining the sodium hydroxide concentration at 150 mM and increasing the sodium acetate concentration during the analysis. The detector response per HCOH group in D-gluco-oligomers (DP 2-7) was almost the same.     

Transfructosylation of rebaudioside A (a sweet glycoside of Stevia leaves) with Microbacterium beta-fructofuranosidase.

It was found that a beta-fructofuranosidase produced by Microbacterium sp. H-1 has potent trans-beta-fructofuranosylation activity from sucrose (donor). By means of this enzyme system, rebaudioside A (RA), the second major sweet steviol glycoside of the leaves of Stevia rebaudiana, was subjected to transfructosylation, affording a mono-beta-fructofuranosylated product (RA-F) in a high yield. The structure of RA-F was elucidated as beta-D-fructofuranosyl-(2----6)-beta-D-glucopyranosyl ester of steviol-13-O-[beta-D-glucopyranosyl-(1----2)]- [beta-D-glucopyranosyl-(1----3)]-beta-D-glucopyranoside. Some improvement in the quality of sweetness was observed for RA-F.     

Saponins from leaves of Acanthopanax hypoleucus Makino.

From the leaves of Acanthopanax hypoleucus Makino (Araliaceae), five triterpenoidal saponins, having oleanolic acid and hederagenin as sapogenins, were isolated. On the basis of chemical and spectral data, the structures of two new saponins, named hypoleucosides A (1), and B (5) were elucidated as follows: 1; 3-O-beta-D-glucopyranosyl 11 alpha-methoxy-oleanolic acid 28-O-beta-D-glucopyranosyl ester, 5; 3-O-beta-D-glucopyranosyl-(1----2)-alpha-L-arabinopyranosyl-(1---- 4)-beta-D-glucopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1----6)-beta-D-glucopyranosyl ester.     

Mitogenic and colony-stimulating factor-inducing activities of polysaccharide fractions from the fruit bodies of Dictyophora indusiata Fisch

Biological effects (mitogenic and colony-stimulating factor (CSF)-inducing activities) of five homogeneous polysaccharides and a conjugated polysaccharide fraction isolated from the fruit bodies of Dictyophora indusiata Fisch. were investigated. Fucomammogalactan (T-3-Ad) and conjugated polysaccharide fraction (T-2-A) exhibited significant mitogenic and CSF-inducing activities. Among two beta-(1----6)-branched (1----3)-beta-D-glucans (T-4-N and T-5-N), only T-4-N showed both mitogenic and CSF-inducing effects. Partially O-acetylated (1----3)-alpha-D-mannans (T-2-HN and T-3-M') did not show these effects.     

Study on the constituents of Desmodium styracifolium

Two triterpenoid saponins (1 and 2) were isolated from Desmodii Herba [the dried whole plants of Desmodium styracifolium (Osbeck) Merr. (Leguminosae)] and their chemical structures were characterized as soyasaponin I and a new saponin, 3-O-[alpha-L-rhamnopyranosyl-(1----2)-beta-D-galactopyranosyl- (1----2)-beta-D-glucuronopyranosyl]soyasapogenol E, respectively, by chemical and spectroscopic means.     

Separation of the hot water extracts of sclerotia of Sclerotinia sclerotiorum IFO 9395 into mitogenic and antitumor-active subfractions

The hot water extract of sclerotia of Sclerotinia sclerotiorum IFO 9395 (TSHW) was divided into representative fractions by ammonium sulfate and ethanol precipitations, and (1----3)-beta-D-glucanase treatment. The ammonium sulfate and ethanol precipitations gave a (1----3)-beta-D-glucan fraction (TSG) and a mannan fraction (TSM). After the degradation of (1----3)-beta-D-glucan in TSHW by (1----3)-beta-D-glucanase treatment, a water-insoluble protein fraction (EDP) and supernatant (EDS) were obtained. Among these fractions, the mitogenic and antitumor activities were mainly observed in EDP and TSG, respectively. On the other hand, the stimulatory effect on the reticuloendothelial system was mainly found in EDP and EDS, and a weak effect was observed in TSG. These findings suggest that the mitogenic and antitumor activities of TSHW were mainly due to the protein and (1----3)-beta-D-glucan, respectively, and that the mitogenic substance (EDP) is tightly bound to (1----3)-beta-D-glucan (TSG) in TSHW, accounting for its solubility in aqueous solution.     

Polysaccharides in fungi. XXVI. Two branched (1----3)-beta-D-glucans from hot water extract of Yu ?r

Two water-insoluble glucans, U-3-N ([alpha]D +1.0 degree, 0.5 M sodium hydroxide) and U-3-AP1 ([alpha]D +2.5 degrees, 1 M sodium hydroxide) were isolated from hot-water extract of the fruiting bodies of Y? ?r (Chinese name) (Auricularia sp.). U-3-N and U-3-AP1 were investigated by a combination of chemical and spectroscopic methods. The results indicated that U-3-N (molecular weight, 6.1 x 10(5)) was similar to beta-(1----6)-branched (1----3)-beta-D-glucan (N-5P: molecular weight, 5.6 x 10(5)) isolated from the alkaline extract of the fruiting bodies, and U-3-AP1 (molecular weight, 6.3 x 10(4)) was beta-(1----6)-branched (1----3)-beta-D-glucan containing beta-(1----6)-linked D-glucopyranosyl residues. U-3-N showed potent anti-tumor activity against the solid form of sarcoma 180, although U-3-AP1 had little effect on the tumor.     

Reticuloendothelial system-activating polysaccharides from rhizomes of Panax japonicus. I. Tochibanan-A and -B

From rhizomes of Panax japonicus (Araliaceae), two polysaccharides named tochibanan-A and -B, which show reticuloendothelial-potentiating activity in the carbon clearance test in mice, were isolated. The structure of tochibanan-A (molecular mass: 23,000) was elucidated as a linear beta-1,4-D-galactan. Tochibanan-B (molecular mass: 40,000) consists of D-galactose (87.1%), L-arabinose, D-glucose and D-galacturonic acid and has a beta-D-(1----4)-linked galactopyranosyl backbone possessing GalA-(1----6)-Gal, Ara-(1----5)-Ara, Gal, and Glc side chains. The structure around the branching points and the repeating unit were investigated and a possible structure of tochibanan-B is proposed.     

Studies on absorption, distribution, excretion and metabolism of ginseng saponins. V. The decomposition products of ginsenoside Rb2 in the large intestine of rats

The decomposition of ginsenoside Rb2 (Rb2) in the rat large intestine after oral administration was investigated in detail. A part of Rb2 was decomposed and six decomposition products (I-VI) were observed on thin-layer chromatogram. Among them, five products (I-V) were isolated, and identification of these compounds was done by carbon-13 nuclear magnetic resonance (13C-NMR). On the basis of 13C-NMR analysis, these compounds were identified as ginsenoside Rd (I), 3-O-beta-D-glucopyranosyl-20-O- [alpha-L-arabinopyranosyl(1----6)-beta-D-glucopyranosyl]-20(S)- proto-panaxadiol (II), ginsenoside F2 (III), 20-O-[alpha-L-arabinopyranosyl(1----6)-beta-D-glucopyranosyl]-20(S )- protopanaxadiol (IV), and compound K (V), respectively.