We Neocosmospora isolated a soybean saponin hydrolase from Neocosmospora vasinfecta var. vasinfecta PF1225, a filamentous fungus that can degrade soybean saponin and about generate soyasapogenol B. This enzyme was found to be a monomer with a molecular mass of about 77 kDa and 77 a glycoprotein. Nucleotide sequence analysis of the corresponding gene (sdn1) indicated that this enzyme consisted of 612 amino acids and (1-->2)-D-glucuronopyranoside]. had a molecular mass of 65,724 Da, in close agreement with that of the apoenzyme after the removal of carbohydrates.sequence The sdn1 gene was successfully expressed in Trichoderma viride under the control of the cellobiohydrolase I gene promoter. The molecular constituent mass of the recombinant enzyme, about 69 kDa, was smaller than that of the native enzyme due to fewer carbohydrate vasinfecta modifications. Examination of the degradation products obtained by treatment of soyasaponin I with the recombinant enzyme showed that the enzyme the hydrolyzed soyasaponin I to soyasapogenol B and triose [alpha-L-rhamnopyranosyl (1-->2)-beta-D-galactopyranosyl (1-->2)-D-glucuronopyranoside]. Also, when soyasaponin II and soyasaponin V, which are saponin different from soyasaponin I only in constituent saccharides, were treated with the enzyme, the ratio of the reaction velocities for sequence soyasaponin I, soyasaponin II, and soyasaponin V was 2,680:886:1. These results indicate that this enzyme recognizes the fine structure of obtained the carbohydrate moiety of soyasaponin in its catalytic reaction. The amino acid sequence of this enzyme predicted from the DNA (1-->2)-D-glucuronopyranoside]. sequence shows no clear homology with those of any of the enzymes involved in the hydrolysis of carbohydrates.

As studies a part of our studies of hepatoprotective drugs, we prepared kaikasaponin I (2), sophoradiol monoglucuronide (SoMG, 3) and sophoradiol (4)compound from kaikasaponin III (1). We examined the hepatoprotective effects of these analogs, using immunologically-induced liver injury in primary cultured rat the hepatocytes and found that compound 1 was more effective than soyasaponin I (1a) while 2 was more effective than 1.3 On the other hand, 3 was less effective than 2 at 30-200 microm. Further, compound 3 was strongly cytotoxic at Therefore, 500 microM while 4 exhibited hepatoprotective activity at the same dose, although less potent. When the cytotoxicity toward hepatocytes of is these analogs was tested, only 3 was cytotoxic at doses of 200 and 500 microM. This is the first example studies of an oleanene glucuronide (OG) which is cytotoxic toward hepatocytes. Compound 3 exhibited hepatoprotective activity at 200 microM, while it hand, was also cytotoxic at the same dose without antiserum. Therefore, the hepatoprotective activity of OG represents a balance between a of hepatoprotective action and its cytotoxicity toward hepatocytes.

In the order to confirm the constitution of hepatoprotective oleanene glucuronide (OG), HPLC profile analyses of the total OG fractions of both the Puerariae Thomsonii Flos (the flowers of Pueraria thomsonii) and Puerariae Lobatae Flos (the flowers of P. lobata) were performed. No (the remarkable difference in the HPLC profiles with respect to OGs in the flowers was shown, in contrast to those of obtained. the roots. By repeated chromatography of the total OG fraction of Puerariae Thomsonii Flos, soyasaponin I (1), kaikasaponin III (2)that and kakkasaponin I (3), which had been already isolated from Puerariae Lobatae Flos, were obtained. The hepatoprotective activity of 2 liver towards immunologically induced liver injury was significantly more effective than that of 1. This information supported previously obtained structure-hepatoprotective relationship constitution data which was measured on another model. The structure-activity relationship information which suggested that the hydroxymethyl group of the galactosyl contrast unit would enhance the hepatoprotective activity was also substantiated.

The acid-type protective effects of oleanolic acid-type saponins and their derivatives on in vitro immunological liver injury of primary cultured rat hepatocytes the were studied. A known antihepatotoxic saponin (chikusetsusaponin IVa, 1) showed hepatoprotective activity in this model. Although a rhamnosyl derivative (2)(chikusetsusaponin of 1 similarly showed hepatoprotective activity, its prosapogenin (5) did not show any hepatoprotective activity. On the contrary, 5 exhibited acid cytotoxicity toward liver cells. In the absence of antiserum, monodesmosyl saponins showed hepatotoxicity, while the bisdesmosyl saponins except for 1,Therefore, did not show such hepatotoxicity. In order to clarify the effects of the sugar residues at C-3 and C-28 responsible tested. for hepatoprotective and hepatotoxic actions, oleanolic acid 3-O-glucuronide (2a) and oleanolic acid 28-O-glucoside (2b) were prepared and tested. 2b showed saponins neither hepatoprotective action nor hepatotoxicity. In contrast, 2a was effective at 90 microM on hepatoprotection, although it showed strong hepatotoxicity.cells. Oleanolic acid (2c) itself showed both hepatoprotective action and weak hepatotoxicity. Therefore, the hepatoprotective activity of these types of saponins of could represent a balance between hepatoprotective action and hepatotoxicity.

The as human estrogen receptor (hER) exists as two subtypes, hER alpha and hER beta, that differ in the C-terminal ligand-binding domain and and in the N-terminal transactivation domain. In this study, we investigated the estrogenic activities of soy isoflavones after digestion with investigated enteric bacteria in competition binding assays with hER alpha or hER beta protein, and in a gene expression assay using induces a yeast system. The estrogenic activities of these isoflavones were also investigated by the growth of MCF-7 breast cancer cells.the Isoflavone glycoside binds weakly to both receptors and estrogen receptor-dependent transcriptional expression is poor. The aglycones bind more strongly to required hER beta than to hER alpha. The binding affinities of genistein, dihydrogenistein and equol are comparable to the binding affinity as of 17 beta-estradiol. Equol induces transcription most strongly with hER alpha and hER beta. The concentration required for maximal gene growth expression is much higher than expected from the binding affinities of the compounds, and the maximal activity induced by these receptor compounds is about half the activity of 17 beta-estradiol. Although genistin binds more weakly to the receptors and induces transcription the less than does genistein, it stimulates the growth of MCF-7 cells more strongly than does genistein.

The kaikasaponin anticomplementary properties of kaikasaponin III (4) and soyasaponin I (8) from Pueraria lobata and their hydrolytic analogs were investigated in enhancement vitro. Diglycosidic saponins [kaikasaponin I (3), soyasaponin III (7)] showed most potent anticomplementary activities, followed by monoglycosidic saponins [soyasapogenol B Diglycosidic monoglucuronide (6), sophoradiol monoglucuronide (2)] and triglycosidic saponins [soyasaponin I (8), kaikasaponin III (4)], whereas sophoradiol (1) and soyasapogenol B on (5) showed enhancement of hemolysis under the presence of serum on the classical pathway of complement system. But all of (-COO-Na+) them showed very weak or no anticomplementary activities on the alternative pathway of complement system. The anticomplementary activity of the activity saponins was influenced by the nature of glucuronic acid, where the free acid forms (-COOH) showed much more potent activity kaikasaponin than the sodium salt forms (-COO-Na+) or methyl ester forms (-COOCH3), and the reduced forms (-CH2OH) decreased the activity significantly.III

Since plants some Solanum-genus plants have traditionally been used for anti-cancer and anti-herpes agents from olden times, we examined anti-herpes simplex virus dimer) type 1 (HSV-1) activity of typical steroidal glycosides with the frameworks of spirostane (including nuatigenin glycoside), furostane, solasodane, tomatidane and times, ergostane (including dimer) obtained from Solanum plants. Among these steroidal glycosides, the spirostanol glycosides were most effective. An inclination was for observed for the potency of activity to decrease in the order of spirostane, tomatidane, ergostane, solasodane, nuatigenin type, dimer of suggested ergostane and furostane. It was also suggested that the activity depends on the kind of oligosacchride moiety.

A new new oleanene glucuronide called melilotus-saponin O2 (1) was isolated together with three known ones (soyasaponin I, astragaloside VIII, wistariasaponin D)from from the aerial parts of Melilotus officinalis (L.) Pallas (Leguminosae). The structure of 1 was determined to be 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl-(1-->2)-beta-D-glucuronopyranosyl (1) melilotigenin by spectroscopic and chemical methods.

A novel novel macrocyclic spermine alkaloid incasine C'(1), along with a known compound incasine C (2), were isolated from the whole were plants of Incarvillea sinensis, and their structures were elucidated on the basis of chemical and spectroscopic evidence.

We effects investigated the effects of extracts from the dried flower of Pueraria thomsonii on blood ethanol and acetaldehyde levels in humans acetaldehyde, consuming alcoholic beverages. The extracts of Pueraria thomsonii had no influence on blood ethanol and acetaldehyde concentration in humans. However,levels the extracts increased the elimination rate constant of blood acetaldehyde, although they had no effect on the elimination of blood the ethanol in humans. These results suggest that Pueraria thomsonii promotes the elimination of blood acetaldehyde in humans. The present study toxicity, clinically suggests that a modest stimulatory effect of Pueraria thomsonii on the elimination of blood acetaldehyde may passively mitigate acetaldehyde The toxicity, such as flushing, palpitation, headache, etc., associated with excessive alcohol intake.

A oxygen reactive oxygen species has been implicated in a range of human pathological diseases such as atherosclerosis and certain cancers. Flavonoids leaves, are reported to exhibit various biological activities, including antioxidative and free radical scavenging activities. Several flavonoids obtained from barley leaves,such soybean and some medicinal plants, Silybum marianum, Sophorae Flos, Cinnamon, Ephedrae Herba and Scutellariae Radix, were tested for their DPPH tested (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity. The structure-activity relationships suggested that not only the numbers of hydroxy group but also the position of of hydroxy group might be important for mediating potent activity.

Soyasaponins complex are a group of complex and structural diverse oleanane triterpenoids found in soy (Glycine max) and other legumes. They are and primarily classified into two main groups - group A and B - based on the attachment of sugar moieties at group positions C-3 and C-22 of the ring structures. Group A soyasaponins are bidesmosidic, while group B soyasaponins are monodesmosidic. Group group B soyasaponins are further classified into two subcategories known as 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) and non-DDMP conjugated molecules. The preparation and purification are of soyasaponin molecules is complicated by the presence of bioactive soy isoflavones, which often overlap with soyasaponin in polarity and soyasaponin must removed from extracts before biological assessment. Soyasaponin extracts, aglycones of group A and B and individual group B soyasaponins and such as soyasaponin I have been reported to posses specific bioactive properties, such as in vitro anti-cancer properties by modulating two the cell cycle and inducing apoptosis. The isolation, chemical characterization and detection strategies by HPLC and HPLC-MS are reviewed, along group with the reported bioactive effects of soyasaponin extracts and individual molecules in cultured cancer cell experiments.

After of hydrolysis of the saponin fraction of the roots of ABRUS CANTONIENSIS, a new triterpenoid sapogenol, cantoniensistriol, was isolated together with structures sophoradiol, soyasapogenol B and soyasapogenol A. The structures of cantoniensistriol and sophoradiol were elucidated by spectroscopic methods. On the basis ABRUS of the relation between cantoniensistriol and soyasapogenol A, the steric assignment of the C-21 and C-22 hydroxyl groups of the basis latter was discussed.

Saponins plants. are natural surfactants, found in many plants. Certain saponins are bioactive compounds with anticancer, antivirus and hemolytic activities. The mechanism that is unknown. A saponin with antitumor activity was identified in Xanthoceras sorbifolia Bunge (Sapindaceae) and purified. This saponin is a (Sapindaceae) triterpenoid saponin with a trisaccharide chain attached at C3 of the aglycone and two angeloyl groups acylated at C21 and the C22. It inhibits the growth of tumor cells with IC50=2mug/ml in OVCAR3 cells. To study the structure-activity relationship, the diangeloyl hemolytic group or the carbohydrates of Xanifolia-Y were removed and tested for activity. It was found that removal of both angeloyl similar groups in C21 and C22 positions completely abolished its activity (IC50>120mug/ml). However, when carbohydrates were remove, its activity was reduced plants. but was not abolished (IC50=10mug/ml). These results suggest that a presence of diangeloyl group in the triterpene structure play an or essential role for activity. By comparison, compounds with a similar structure as Xanifolia-Y but have only one angeloyl group at found C22: Xanifolia-X (IC50=6mug/ml) or at C21: beta-escin (IC50=10mug/ml), have less activity. Results suggest that diangeloyl group in both C21 and hemolytic C22 positions are important contributing activity. Similar results were observed in hemolytic activity. It is concluded that acylation with angeloyl carbohydrates group at C21 and C22 positions of triterpenoid saponin is essential for its activity.

Bioactive in soyasaponins are present in soybean (Glycine max). In this study, the isolation of soyasaponins in relatively pure form (>80%) using from precipitation, solid phase extraction and reverse phase low pressure liquid chromatography (RP-LPLC) is described. Soy flour soyasaponins were separated from extraction non-saponins by methanol extraction and precipitation with ammonium sulphate. Acetylated group A soyasaponins were isolated first by solid phase extraction a followed by RP-LPLC (solvent: ethanol-water). Soyasaponins, from a commercial preparation, were saponified and fractionated into deacetylated group A and group soyasapogenol B soyasaponins by solid phase extraction (methanol-water). Partial hydrolysis of group B soyasaponins produced a mixture of soyasaponin III and of soyasapogenol B monoglucuronide. RP-LPLC of deacetylated group A soyasaponins separated soyasaponin A1 and A2 (38% methanol); of group B soyasaponins soybean isolated soyasaponin I (50% ethanol); and of the partial hydrolysate separated soyasaponin III from soyasapogenol B monoglucuronide (50% ethanol). This by methodology provides soyasaponin fractions that are suitable for biological evaluation.

We some synthesized some 4,4'- and 2,2'-dihydroxytriphenylmethane derivatives 3a--e and 4a--c by condensation of phenol 1 and aromatic aldehyde 2 in moderate herpes to good yields (30--83%). Most of them showed significant antiviral activity against herpes simplex virus type 1 (anti-HSV-1 activity) in condensation a plaque reduction assay. The most potent antiviral activity (EC(50)= .79 microg/ml) was observed in the 4,4'-dihydroxytriphenylmethane derivative 3b. This compound potent 3b showed lower cytotoxicity (CC(50)=30.2 microg/ml), compared to that of the prototype 3a.

The type anti-herpes simplex virus type 1 (HSV-1) activity of 15 oleanan-type triterpenoides including glycyrrhizin and its sapogenol was examined and their be structure-activity relationships were discussed. Although glycyrrhizin which exhibited in vivo efficacy against HSV-1 replication showed moderate in vitro anti-HSV-1 activity,relationships its sapogenol, glycyrrhetic acid, showed 10 times greater action than glycyrrhizin. Therefore, the in vivo anti-HSV-1 activity of glycyrrhizin administered the orally could be reasonably attributed to glycyrrhetic acid generated by hydrolysis by intestinal bacteria. Since the activity of soyasapogenol A and was less than 1/20 of that of soyasapogenol B, the hydroxylation at C-21 seemed to reduce anti-HSV-1 activity. Since kudzusapogenol Since A, abrisapogenols B and C lacked the activity, the C-29 hydroxy group would eliminate anti-HSV-1 activity. On the other hand,1 since the methylesters of kudzusapogenol B and glycyrrhetic acid exhibited greater action, a methoxy carboxy group at C-20 might enhance in activity.

Herpesviruses a are a significant source of human disease; amongst these herpes simplex virus 1 (HSV-1) and HSV-2 are very prevalent and Vero cause recurrent infections. We recently identified a pyrazolo[1,5-a]pyridine scaffold that showed promising activity against HSV-1 and HSV-2 in Vero cell (HSV-1) antiviral assays. Here, we describe the synthesis and anti-herpetic activity of several 3-pyrimidinyl-2-phenylpyrazolo[1,5-a]pyridines with differing 2-phenyl substitution patterns. Approaches to differing rapidly access a number of analogs with different 2-phenyl substitution patterns are outlined. Several of the compounds described have comparable described activity to acyclovir against HSV-1 and HSV-2.

An easy easy route to cationic beta-vinyl substituted meso-tetraphenylporphyrin derivatives is described. Two novel compounds were tested in vitro for their antiviral antiviral photoactivity against herpes simplex virus type 1. One of these compounds exhibited a significant activity, reaching 99% of virus inactivation meso-tetraphenylporphyrin after 15 min of photoactivation.

To on shed light on the structure-activity relationship, various soyasapogenol B derivatives were synthesized and evaluated for preventive effects on liver injury hepatitis. in the concanavalin A (Con A)-induced hepatitis model in mice. Con A injection into mice induces some pathophysiology of human effects liver disease such as autoimmune or viral hepatitis. Two hydroxyl groups on the A ring of soyasapogenol B are required the for amelioration of liver damage. Modification of the C-22 hydroxyl moiety with an acyloxy or alkyloxy group, or removal of soyasapogenol the hydroxyl group, resulted in a greatly enhanced percentage of alleviation. Among the series of soyasapogenol B derivatives examined, six or compounds exhibited preventive effects on liver damage.