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Based on NOE experiments, the structure of naphthodipyranodione from Gentianaceae plants was revised to 1,2-dihydro-4H,6H,8H-naphto[1,2-d:4,5- c'd']dipyrano-4,8-dione. Naphthodipyranodione was assumed to be formed by the degradation of gentiopicroside by enzymatic hydrolysis at low water -concentration. The degradation pathway was a unique domino-reaction triggered by enzymatic hydrolysis. Naphthodipyranodione may become an index compound for the drying and/or fermenting procedure of Gentian root.

Achyranthoside H methyl ester (AH-Me) is an oleanolic acid saponin derivative isolated from the roots of Achyranthes fauriei through diazomethane treatment. AH-Me exhibited significant cytotoxicity against human breast cancer MCF-7 and MDA-MB-453 cells, with respective ID(50) values of 4.0 and 6.5 muM: in the MTT assay. AH-Me is a unique saponin containing three methoxycarbonyl groups in the sugar moiety linked to the C-3 position of oleanolic acid. The demethylation of these methoxycarbonyl groups by alkaline hydrolysis caused a marked reduction of the cytotoxicity of AH-Me, suggesting that the methoxycarbonyl groups of AH-Me are key groups for the acquisition of cytotoxicity against human cancer cells. The staining of cancer cells with 4',6'-diamidino-2-phenylindole (DAPI) showed that the population of cells with altered nuclear morphology, for example chromatin condensation and fragmentation, increased markedly after AH-Me treatment. Exposure of MCF-7 and MDA-MB-453 cells to AH-Me resulted in a dose-dependent and time-dependent increase in the sub-G1 population, and in the cleavage of poly-ADP-ribose polymerase (PARP) followed by the formation of an 89 kD peptide. Pretreatment of the cells with the pan-caspase inhibitor z-VAD-fmk abolished the cleavage of PARP by AH-Me treatment and suppressed the antiproliferative effect of AH-Me on tumor cell growth. These results together led to the suggestion that AH-Me induces apoptosis via the caspase activation pathway in human breast cancer cells, and apoptosis is the major mode of the cytotoxic effect triggered by AH-Me.

Two oleanolic acid saponins named achyranthosides G (1) and H (2) were newly isolated from Achyranthes fauriei root as methyl esters in addition to methyl esters of achyranthosides A - F and five oleanolic acid glucuronides (chikusetsusaponins IVa, V, 28-deglucosyl chikusetsusaponin V, pseudoginsenoside RT(1), and oleanolic acid 3-O-beta-D-glucuronopyranoside) as well as oleanolic acid 28-O-beta-D-glucopyranoside, beta-ecdysterone, and polypodine B. Their structures were characterized as follows on the basis of the chemical and spectroscopic evidences.

Amberlite XAD-7, a hydrophobic polymer, was used to change microbial reaction of ketones from reduction to Baeyer-Villiger (BV) oxidation. Thus, D. magnusii NBRC 4600 and G. reessii NBRC 1112 could catalyze the BV reaction of ketones in the presence of the polymer while reduction of the substrates proceeded, and BV oxidation was scarcely found in the absence of the polymer.

We isolated and characterized 23 compounds, including a new iridoid named gentiolutelin and its dimethyl acetal, and a new lignan named gentioluteol from fresh roots (including small amounts of rhizome) of Gentiana lutea cultivated in Japan (Hokkaido). The structures of gentiolutelin and gentioluteol were determined as (1S,2R,3S,5R)-3-hydroxy-2-methyl-5-(2-oxoethyl)-cyclopentanecarboxylic acid methyl ester and (7R,8S,8′S)-4,4′,8,9-tetrahydroxy-3,3′,5-trimethoxy-7,9′-epoxylignan, respectively, on the basis of chemical and spectroscopic evidence. It was noteworthy that gentiopicroside, known to be a major secoiridoid glycoside in Gentian root, was not detected from the fresh roots of 3-year-old G. lutea.

N-para-ferrocenyl benzoyl dipeptide esters 2–5 were prepared by coupling para-ferrocenyl benzoic acid 1 to the dipeptide ethyl esters GlyGly(OEt) 2, GlyAla(OEt) 3, GlyLeu(OEt) 4 and GlyPhe(OEt) 5. The compounds were fully characterized by a range of NMR spectroscopic techniques, mass spectrometry (ESI-MS, FABMS, MS/MS) and cyclic voltammetry (CV). The a1 and b1 ions appeared at one mass unit less than the expected mass in the tandem mass spectra. The CV curves of compounds 2–5 exhibited quasi-reversible redox behavior similar to the Fc/Fc+ redox couple and displayed oxidation potential values in the 50–55 mV range, against the Fc/Fc+ redox couple.

rac-Hept-1-en-3-ol 4 was subjected to an enantioselective esterification in the presence of Novozyme 435 and vinyl crotonate as the acyl donor to give (3S)-oct-1-en-3-yl crotonate 7 in >99% ee and (3R)-alcohol 4 in 99% ee. The E-value of this enzymatic reaction was found to be >1000. The (S)-crotonic ester 7 was converted by ring-closing metathesis (RCM) using Grubbs’ catalyst to give (S)-oct-2-en-4-olide 1 in 96% yield while keeping the high enantiomeric excess.

Chiral enolate derived from (4R)-4-tert-butyldiphenylsilyloxymethyl-4-butanolide 10 with lithium hexamethyldisilyazide (LiHMDS) was treated with trisyl azide, followed by addition of TMSCl to give (2S,4R)-2-azido-4-[(tert-butyldiphenylsilyloxy)methyl]-4-butanolide 8 (53%), from which the first total synthesis of polyoxin M (1) was achieved in overall 3.2% yield (13 steps) from d -glutamic acid. Moreover, the synthesis of the reported synthetic intermediate (2S,4R)-4-hydroxyornithine congener 6 for biphenomycins A and B was also achieved in overall 4.1% yield (12 steps) from d -glutamic acid.

Direct beta-glucosidation between (-)-myrtenol and nerol and D-glucose (3) using the immobilized beta-glucosidase from almonds with the synthetic prepolymer ENTP-4000 gave myrtenyl O-beta-D-glucoside (4) and neryl O-beta-D-glucoside (10), respectively. The coupling of the myrtenyl or neryl O-beta-D-glucopyranoside congeners (7 or 13) and 2,3,4-tri-O-benzoyl-beta-L-arabinopyranosyl bromide (8) afforded the coupled products (9 or 14), respectively. Deprotection of the coupled products (9 or 14) afforded the synthetic myrtenyl 6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside (Sacranoside A, 1) or neryl 6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside (Sacranoside B, 2), respectively.

Three known saponins, chiisanoside, 11-deoxyisochiisanoside, and isochiisanoside, and one novel saponin, 3,4-seco-4(23),20(29)-lupadiene-3,28-dioic acid 28-O-alpha-l-rhamnopyranosyl (1-->4)-beta-d-glucopyranosyl (1-->6)-beta-d-glucopyranoside, referred to as sessiloside, were isolated from a hot water extract of Acanthopanax sessiliflorus leaves. All of these saponins were lupane-type triterpene triglycosides, and their concentrations were 4.1, 1.0, 0.5, and 0.4% (w/w) of the total extract, respectively. Sessiloside and chiisanoside inhibited pancreatic lipase activity in vitro, and addition of the saponin-rich fraction to a high-fat diet suppressed the body weight gain of mice. The possibility of application of the lupane-type saponins from A. sessiliflorus leaves to the treatment of obesity is discussed.

Gentianae Radix, the dried root and rhizoma of Gentiana lutea L. (Gentianaceae), has long been used as a remedy for liver and stomach inflammation, eye troubles, etc. In this paper, the gastroprotective effects of the methanol extract of Gentian root (GM) were studied using different gastric lesion models. In pylorus-ligated rats, administration of GM in the duodenum suppressed gastric juice secretion and total acid output in a dose-dependent manner. Oral or duodenum administration of GM showed significant protection against acute gastric ulcer induced by aspirin plus pylorus ligation, water-immersion restraint stress-induced ulcers, and gastric mucosal injury induced by ethanol. Furthermore, four secoiridoid glycosides, amarogentin (A1), gentiopicroside (A2), amaroswerin (A3), and swertiamarin (A4), were obtained from Gentian root or Swertia herb, and their protective effects against stress-induced ulcers and ethanol-induced gastric mucosal injury were evaluated. The doses required for 50% inhibition (ID50) of A1, A3, and A4 on stress-induced ulcers were calculated to be 5.76, 2.58, and 167 mg/kg respectively. The protective effect of A2 at 250 mg/kg was 26.5%. On ethanol-induced gastritis, 5.0 mg/kg of A1 and A3 showed remarkable suppressive effects (33.7 and 45.4%, respectively), and 20 mg/kg of A4 exhibited a suppressive effect (30.8%). The effects of A1, A3, and A4 on ethanol-induced gastric lesions were canceled by 5.0 mg/kg indomethacin pretreatment. These results suggest that the therapeutic effects of Gentian root on gastric lesions are associated with enhanced mucosal defensive factors via the prostaglandin pathway in the cell membrane, and that secoiridoid glycosides contribute to this activity.

In Japan, pharmacists' activities for the most part consist of dispensing although in some University Hospitals they are directly involved in patient care. In the United States, pharmacists' activities have evolved over forty years in providing drug therapy and have now expanded to improvement in the patient's quality of life. In addition, a six-year pharmacy education program based on patient care is now in place nationally. Furthermore, World Health Organization (WHO) and International Pharmaceutical Federation (FIP) have made recommendations on pharmacists' activities. Shifting to a six-year pharmacy education in Japan has now been decided, and new approaches are being proposed. For pharmacists to serve society in their role as health care professionals, one needs to examine the activities they are expected to perform and pharmacy education necessary to develop these skills. In this paper, pharmacy education was examined by analyzing and comparing Western countries and Japan, with a focus on Canadian pharmacists' activities and pharmacy education in Alberta.

Direct beta-glucosidation between benzyl alcohol and D-glucose (5) using the immobilized beta-glucosidase from almonds with the synthetic prepolymer ENTP-4000 gave a benzyl beta-D-glucoside (1) in 53% yield. The coupling of the benzyl beta-D-glucopyranoside congener (8) derived from 1 with phenyl 2,3,4-tri-O-acetyl-1-thio-beta-D-xylopyranoside (9), ethyl 2,3,4-tri-O-acetyl-1-thio-alpha-L-rhamnopyranoside (13), and 2,3,4-tri-O-acetyl-alpha-L-arabinopyranosyl bromide (15) afforded 10, 14, and 16, respectively, as coupled products. Deprotection of 10, 14, and 16 provided the synthetic benzyl beta-D-xylopyranosyl-(1--6)-beta-D-glucopyranoside (2), benzyl alpha-L-rhamnopyranosyl-(1--6)-beta-D-glucopyranoside (3), and benzyl alpha-L-arabinopyranosyl-(1--6)-beta-D-glucopyranoside (4), respectively.

A series of N-para-ferrocenyl benzoyl amino acid ethyl esters 1–8 have been prepared by coupling para-ferrocenyl benzoic acid with the amino acid esters using the conventional 1,3-dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt) protocol. The amino acids employed in the synthesis were glycine, l -alanine, l -leucine, l -phenylalanine, β-alanine, 4-aminobutyric acid, (±)-2-aminobutyric acid and 2-aminoisobutyric acid. The compounds were fully characterized by a range of spectroscopic techniques such as NMR and mass spectrometry. In addition the X-ray crystal structures of the glycyl 1 and (±)-2-aminobutyrate 7 derivatives have been determined. Analysis of relevant fragments in crystal structures on the Cambridge Structural Database indicates a relative paucity of common fragments such as the α-aminobutyrate group in comparison to the glycyl moiety.

The N-ferrocenoyl amino acid ester derivatives FcCOR {Fc=(η5-C5H5)Fe(η5-C5H4)} where R=Gly(OMe) 1, Gly(OEt) 2, Gly(OBn) 3, l -Ala(OMe) 4, l -Ala(OEt) 5, l -Leu(OMe) 6, l -Leu(OEt) 7, l -Leu(OBn) 8, l -Phe(OMe) 9 and l -Phe(OEt) 10, were prepared by coupling ferrocene carboxylic acid with the appropriate amino acid ester starting materials using the 1,3-dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt) protocol and these have been characterised by spectroscopic techniques. The electrochemical anion sensing behaviour of compounds 1–10 with several anions using a platinum microdisk working electrode is described, together with 1H NMR anion complexation studies. The X-ray single crystal structure of N-ferrocenoyl- l -alanine methyl ester 4 has been determined and contains two molecules which differ slightly in conformation in the asymmetric unit of space group P21 (No. 4); principal dimensions are amide N(H)CO 1.224(6) and 1.231(6) A, ester CO 1.220(10) and 1.190(7) A, with N–H⋯OC(amide) as the primary intermolecular hydrogen bond, N⋯O 2.992(6) and 2.971(6) A and with graph set C(4).

For the purpose of finding the best reaction conditions of β-glucosidation of primary alcohols, screening experiments in respect of the enzymes, glycosyl donors and solvents were carried out. The effective enzyme and glycosyl donor for the synthesis of benzyl β- d -glucopyranoside (4) appeared to be β-glucosidase (EC 3.2.1.21) from almonds and 4-nitrophenyl β- d -glucopyranoside (7), respectively. Enzymatic glycosidation of 10 kinds of primary alcohols (methanol, ethanol, n-propanol, n-butanol, n-pentanol, n-hexanol, 2-methoxyethanol, 2-ethoxyethanol, benzyl alcohol, (S)-(−)-perillyl alcohol) gave stereoselectively the corresponding β- d -glucopyranosides (14–21, 4 and 22) in moderate yield, respectively.

We here show that single-stranded oligonucleotides containing 5-formyl-2'-deoxyuridine (fdU) can crosslink the peptides derived from the DNA binding site of RecA protein through a Schiff base formation. The ability of crosslinking of fdU-containing oligonucleotides was investigated using a series of peptides whose amino acid residues spanning the center of the RecA-derived peptide were sequentially replaced with lysine. Circular dichroism (CD) spectroscopy, gel mobility shift assay and sedimentation experiment demonstrated that crosslinking reaction proceeded efficiently only when the peptides bound to the oligonucleotides.

The present study was undertaken to investigate the acute anti-fatigue effect of a liquid nutritive and tonic crude drugs (NTDs) on stress induced in mice. After forced walking for 3 or 6h, the NTDs (applied orally, 10 ml/kg) significantly increased locomotor activity, while the administration of NTDs after rapid eye movement (REM) sleep deprivation stress and after immobilization stress did not show a specific effect, having a similar effect as the vehicle with added vitamins, taurine and caffeine. The administration of NTDs after freezing due to electric shock stress showed a specific effect which was not seen in other control groups, water, vehicle (ethanol) and vehicle including vitamins, taurine and caffeine and so resemble the specific effect of NTDs in the stress of forced walking. The present results indicate that the NTDs produced an anti-fatigue effect on the decreased locomotor activity after forced walking and immobility induced by electric stimulation. However, the crude drugs were not effective in improving immobility after sleep deprivation or immobilization stress.

Five sulfonated compounds, namely 4-gingesulfonic acid and shogasulfonic acids A, B, C and D, were isolated together with seven known compounds including 6-gingesulfonic acid from Zingiberis rhizome (Japanese name: Shokyo) made out of ginger. Their structures were characterized by means of spectroscopic analysis.

Screening using a reporter under the control of the hypoxia-response element (HRE) identified several flavonoids and homoisoflavonoids that inhibit the activation of HRE under hypoxic conditions. Among various compounds, isorhamnetin, luteolin, quercetin, and methyl ophiopogonanone B (MOB) were effective at 3 to 9 microg/ml in inhibiting the reporter activity. The expression of vascular endothelial growth factor (VEGF) mRNA during hypoxia was also inhibited by MOB in HepG2 cells, but the effective doses were 10 to 20 microg/ml. MOB caused destabilization of hypoxia-inducible factor (HIF)-1alpha, as revealed by Western blotting, that was dependent on proteasome activity and the tumor suppressor, p53. The tubular formation and migration of human umbilical vein endothelial cells was also inhibited by MOB. MOB is expected to act as an inhibitor of angiogenesis.

From the water-soluble portion of the methanol extract of the fresh rhizome of Atractylodes japonica, five new sesquiterpenoid glycosides, including a compound having a secoguaiane skeleton, and a new aromatic compound glycoside were isolated together with ten known compounds. Their structures were clarified by spectral investigation.

Five sesquiterpenoid glycosides (two guaiane-type glycosides and three eudesmane-type glucosides) and a glucoside of an acetylene derivative were newly isolated from the water-soluble portion of the methanolic extract of Atractylodes lancea rhizome together with 26 known compounds. Their structures were characterized as atractyloside A 14-O-beta-D-fructofuranoside, (1S,4S,5S,7R,10S)-10,11,14-trihydroxyguai-3-one 11-O-beta-D-glucopyranoside, (5R,7R,10S)-isopterocarpolone beta-D-glucopyranoside, cis-atractyloside I, (2R,3R,5R,7R,10S)-atractyloside G 2-O-beta-D-glucopyranoside, and (2E,8E)-2,8-decadiene-4,6-diyne-1,10-diol 1-O-beta-D-glucopyranoside on the basis of chemical and spectroscopic investigation. The presence of six characteristic guaiane-type glucosides in both rhizomes of A. lancea and Atractylodes japonica suggested a close chemotaxonomic relationship between them.

A series of N-para-(ferrocenyl)benzoyl amino-acid esters, para-Fc(C6H4)CONHCH(R)CO2CH3 {Fc = (η5-C5H5)Fe(η5-C5H4); R = H, CH3, CH2CH(CH3)2, CH2C6H5}, 3–6 have been prepared by coupling para-(ferrocenyl)benzoic acid to the amino-acid esters (gly, l -Ala, l -Leu, l -Phe) using the standard 1,3-dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt) protocol. The compounds were fully characterized by a range of spectroscopic techniques including FAB-MS. The X-ray crystal structures of the parent para-(ferrocenyl)benzoyl methyl ester, Fc-C6H4CO2Me, 1 and a chiral derivative N-{para-(ferrocenyl)benzoyl}- l -alanine methyl ester, Fc-C6H4CONHCH(CH3)CO2Me, 4 have been determined.

Screening of various natural products in a search for novel inducers of apoptosis in human leukemia cells led us to identify the strong apoptosis-inducing activity in a fraction extracted with methanol from the roots of Sophora subprostrata Chun et T. Chen. We purified the compound that induced apoptosis in human leukemia cells and identified it as sophoranone. Sophoranone inhibited cell growth and induced apoptosis in various lines of cells from human solid tumors, with 50% inhibition of growth of human stomach cancer MKN7 cells at 1.2 +/- 0.3 microM. The growth-inhibitory and apoptosis-inducing activities of sophoranone for leukemia U937 cells were very much stronger than those of other flavonoids, such as daidzein, genistein and quercetin. At the early stages of treatment of U937 cells with sophoranone, reactive oxygen species were formed, mitochondrial permeability pores were opened and cytochrome c was released from mitochondria. Cytochrome c was also released upon treatment of isolated mitochondria with sophoranone. Inhibitors of complexes III and IV, but not complexes I and II, of the mitochondrial respiratory chain prevented the release of cytochrome c from isolated mitochondria by sophoranone, as well as the induction of apoptosis in U937 cells in response to sophoranone. Our results indicate that sophoranone might be a unique apoptosis-inducing anticancer agent that targets mitochondria.

Based on NOE experiments, the structure of naphthodipyranodione from Gentianaceae plants was revised to 1,2-dihydro-4 H,6 H,8 H-naphto[1,2-d:4,5-c'd']dipyrano-4,8-dione. Naphthodipyranodione was assumed to be formed by the degradation of gentiopicroside by enzymatic hydrolysis at low water concentration. The degradation pathway was a unique domino-reaction triggered by enzymatic hydrolysis. Naphthodipyranodione may become an index compound for the drying and/or fermenting procedure of Gentian root.

Aromatic and aliphatic racemic alcohols were converted to the corresponding optically active alcohols in high yield with excellent enantioselectivities by the use of Geotrichum candidum IFO 5767.

A hydrophobic polymer, Amberlite™ XAD, was used as material to control the stereochemical course of microbial reductions. In the presence of XAD, simple aliphatic and aromatic ketones were reduced to the corresponding (S)-alcohols in excellent enantiomeric excess (ee) while low enantioselectivities were observed in the absence of the polymer.

Five new triterpenoid saponins, nipponosides A-E (1, 3-6), were isolated from Acanthopanax nipponicus leaves, along with a known saponin, kalopanaxsaponin G (2). Nipponosides A-E were characterized as the 28-O-alpha-L-rhamnopyranosyl(1--4)-beta-D-glucopyranosyl(1--6)-beta -D-glucopyranosyl ester of 3-oxohederagenin, oleanolic acid 3-O-beta-D-glucopyranoside, gypsogenin 3-O-beta-D-glucopyranoside, 3beta,23,29-trihydroxyolean-12-en-28-oic acid, and 3beta,20alpha, 23-trihydroxy-30-nor-olean-12-en-28-oic acid, respectively. The structures of these new compounds were based on chemical and spectral methods.

Three monoterpenoid alcohols (one new fenchane-type and two new acyclic) and four monoterpenoid glycosides (two new thujane-type, one camphane-type and one new norfenchane-typ) were isolated from the watersoluble portion of the methanolic extract of fennel. Their structures were clarified by spectral investigation.

From the water-soluble portion of the methanolic extract of fennel, seven new sugar alcohols (two deoxybutitols, four deoxypentitols and one deoxyhexitol) and a sugar lactone were isolated, together with seven known glucides, four nucleosides, (3R)-2-hydroxymethylbutane-1, 2, 3, 4-tetrol and uracil. From the results of spectral investigation, the new compounds were characterized as 1-deoxythreitol (9), (2R)-butane-1, 2, 4-triol (10), 1-deoxy-D-ribitol (11), 1-deoxy-D-xylitol (12), 2-deoxy-D-ribitol (13), 3-deoxyarabinitol (14), 1-deoxy-D-glucitol (16) and 2-deoxy-D-ribono-1, 4-lactone (15), respectively.

beta-Hydroxyisovalerylshikonin (beta-HIVS), which was isolated from the plant, Lithospermium radix, inhibited the growth of various lines of cancer cells derived from human solid tumors at low concentrations between 10(-8) and 10(-6) M. When HL-60 cells were treated with 10(-6) M beta-HIVS for 3 h, characteristic features of apoptosis, such as DNA fragmentation, nuclear fragmentation, and activation of caspase-3-like activity, were observed. The most characteristic features of the effect of beta-HIVS were the remarkable morphological changes induced upon treatment of HL-60 cells with beta-HIVS, as visualized on the staining of actin filaments with phalloidin labeled with tetramethylrhodamine B isothiocyanate. Moreover, activation of MAP kinases, such as ERK2, JNK and p38, was detected after treatment with 10(-6) M beta-HIVS preceding the appearance of the characteristics of apoptosis, and the features of the activation of these MAP kinases were quite different from those of Fas and anticancer drug-induced apoptosis. The activation of JNK by beta-HIVS was not inhibited by inhibitors of caspases, suggesting that JNK is located either upstream or independent of the caspase signaling pathway. beta-HIVS did not inhibit the activity of topoisomerase II. These results indicate that beta-HIVS induces apoptosis in HL-60 cells through a mechanism unlike those reported for anti-Fas antibodies and etoposide.

The interaction between sodium ascorbate and dopamine was investigated by three different parameters: radical intensity, prooxidant action, and cytotoxicity induction. Sodium ascorbate and dopamine produced the doublet and quartet ESR signals under alkaline conditions (pH 8.0-9.5), respectively. Addition of increasing concentrations of sodium ascorbate completely scavenged the dopamine radical and replaced the latter with its own radical. Similarly, dopamine slightly, but significantly reduced the radical intensity of sodium ascorbate. These two compounds stimulated the methionine oxidation and hydrogen peroxide generation in culture medium, but in combination, their stimulation activities were weakened. Both of these two compounds dose-dependently reduced the viable cell number of human oral squamous carcinoma HSC-4 cells, and their cytotoxic activity was significantly reduced by catalase. When these two compounds were mixed together before adding to HSC-4 cells, both of their cytotoxic activities were diminished. The present study demonstrates the interaction between sodium ascorbate and dopamine, which might modify their biological activities and generation of nerve disorders such as Parkinson's disease.

A peptidic mimetic of RNA having a guanine-adenine-guanine code as a base sequence was prepared as a leading compound of Vero-toxin inhibitors. The synthesized hexapeptide is composed from two γ-guanyl-β-hydroxy-α- l -amino acids and one γ-adenyl-β-hydroxy-α- l -amino acid, which were prepared by using l -threonine aldolasecatalyzed reaction, in addition to three glycines. On designing this peptide, the β-hydroxyl groups in the novel α-amino acids are compared to the 2′-hydroxyl groups of RNA, and glycine was chosen as the mimic of the phosphate groups in the RNA backbone.

We synthesized the sialylphosphatidylethanolamine (sialyl PE) derivatives Neu5Ac-PE, (Neu5Ac)2-PE, Neu5Ac-PE (amide) and Neu5Ac-PE (methyl). We examined the anti-viral effects of the derivatives on human influenza A virus infection by ELISA/virus-binding, hemagglutination inhibition, hemolysis inhibition and neutralization assays. The sialyl PE derivatives that we examined bound to A/Aichi/2/68, A/Singapore/1/57 and A/Memphis/1/71 strains of H3N2 subtype, but not to A/PR/8/34 strain of H1N1 subtype. The derivatives inhibited viral hemagglutination and hemolysis of human erythrocytes with A/Aichi/2/68 and A/Singapore/1/57 (H3N2), but not with A/PR/8/34 (H1N1). The inhibitory activity of the (Neu5Ac)2-PE derivative was the strongest of all sialyl PE derivatives (IC50, 35 μM to 40 μM). Sialyl PE derivatives also inhibited the infection of A/Aichi/2/68 in MDCK cells. Complete inhibition was observed at a concentration between 0.3 to 1.3 mM. IC50 of (Neu5Ac)2-PE was 15 μM in A/Aichi/2/68 strain. Taken together, the synthetic sialyl PE derivatives may be effective reagents against infection of some types of influenza A viruses.

Our previous report stated that kikyo-to, a Japanese herbal medicine, consisting of the roots of Platycodon grandiflorum and Glycyrrhiza sp., stimulates the pancreatic exocrine secretion of conscious rats. The present study focused on the effective components of kikyo-to and the mechanism of stimuli to pancreatic secretion of rats. When 10 to 100 mg of platycodin D, a saponin from the root of Platycodon grandiflorum, was intragastrically administered, the pancreatic secretion of rats was stimulated. At the same time, the plasma CCK concentration increased. On the other hand, the stimulative effects of glycyrrhizin, a saponin from the root of Glycyrrhiza sp. were weak compared to platycodin D. The effects of 10 mg/kg of platycodin D on pancreatic secretion were inhibited by loxiglumide (50 mg/kg, i.g.), a CCK receptor antagonist. In contrast, the suppressive effect of atropine (300 micrograms/kg/h, i.v.) on pancreatic secretion was reduced by administering 10 mg/kg of platycodin D. In addition, up to 1 mM of platycodin D did not inhibit the trypsin activities in vitro. In conclusion, kikyo-to serves to stimulate pancreatic exocrine secretion mainly because platycodin D causes gastrointestinal hormones, particularly, CCK to be released from the duodenum.

Further investigation of the leaves of Acanthopanax divaricatus gave two analogues of chiisanoside, which is a lupane triterpenoid oligoglycosyl ester. The structures were established as 28- O -α- l -rhamnopyranosyl(1 → 4)-β- d -glucopyranosyl(1 → 6)β- d -glucopyranosyl esters of 1β,11α-dihydroxy-3-oxo-lup-20(29)-en-28-oic acid and 1( R ),11α,22α-trihydroxy-3,4-seco-lupa-4(23),20(29)-diene-3,28-dioic acid 3,11α-lactone based on chemical and spectroscopic evidence. in biosynthetic terms, one is the precursor of chiisanoside and the other is an oxygenated derivative chiisanoside.

Carbohydrates on cell surfaces play important roles in various cell adhesion processes such as bacterial and viral infection, neurtrophile recruitment, cell differentiation and so on by binding proteins like lectins, antibodies, and glycoenzymes (glycosidases and glycosyltransferases). Carbohydrate analogues which inhibit the interaction between carbohydrates and proteins are promising molecules to clarify and control biological functions of oligosaccharides, and some of the analoges were prepared by using aldolase catalyzed reactions without any tedious procedures. For example, a mimetic of sialyl Lewis X which is an ligand of E-selectin was synthesized by using L-threonine aldolase catalyzed reaction as a key step. In addition, synthetic approach to KDO and 4-azaiduronic acid by using kynureninase and the L-threonine aldolase, respectively, is almost achieved.This review article shows solid phase synthesis and combinatorial synthesis of carbohydrates and treats the details about the utility of aldolases on making carbohydrate mimetics as one of the chemical approaches of glycotechnology.

細胞表層に存在する複合糖鎖の生物学的機能は高次構造を有する糖鎖に抗体,酵素,レクチンなどの糖認識タンパクが結合することによって発現する.E-セレクチンのリガンドとなるシアロルイスXを中心に,エネルギー的に安定な構造と生理活性を発現する構造の違いならびに糖鎖類似体の設計について概説する.

We found that an impurity was formed in the 1% dibucaine injection solution following ordinal treatment using the high-pressure steam method. The impurity was isolated and characterized to be 2-hydroxy-N-[2-(diethylamino) ethy1-4-quinolinecarboxamine (debutyl-dibucaine)], which was caused from the thermal degradation of dibucaine brought about by the sterilization process. The degradation of dibucaine with dealkylation was found to be affected under the thermal treatment, time and temperature conditions. In addition, suitable sterilization conditions were established to maintain the purity of the 1% dibucaine injection solution at more than 99.5%.

Two new saponins, achyranthosides A and B, were isolated from Achyranthes fauriei root, and their structures were elucidated on the basis of chemical and physical evidences. Achyranthoside A methyl ester was found to have significant cytotoxic activity against human colon carcinoma and murine melanoma cells.

From fresh rhizome of Alisma orientale, and the crude drug Alismatis rhizoma of Japanese and Chinese origins, 15 new terpenoids were isolated together with 11 known terpenoids and were characterized as follows: 11-deoxy, 13β,17β-epoxy and ll-deoxy-13β,17β-epoxy derivatives of alisols, alisol C, 16,23-oxidoalisol B, 25-O-methyl and 25-anhydro derivatives of alisol A, as well as eudesma-4(14)-en-1,6-diol, 10-O-methyl-alismoxide and 16(R)-(−)-kaurane-2,12-dione.

Clycosides of (±)-5,5′-dimethoxylariciresinol, 2(p-hydroxy-phenyl)-ethanol and N-methylhigenamine were isolated from the dried bark of Phellodendron amurense, together with nine phenolic compounds.

The hot water extract of adzuki (HWEA), which is produced as a byproduct in the adzuki bean boiling process, has anti-tumor, antioxidative, and anti-diabetic activities. In this study, we fractionated HWEA to 4 fractions using stepwise gradient column chromatography with water and ethanol, and demonstrated the effects of each fraction on antigen (Ag)-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells. The 40% ethanol eluate extract (EtEx.40) showed the strongest inhibition level of these fractions. To reveal the inhibitory mechanisms underlying degranulation by EtEx.40, we investigated intracellular reactive oxygen species (ROS) production, intracellular free Ca²⁺ concentration ([Ca²⁺]i), and early intracellular signaling pathways. Treatment with EtEx.40 markedly inactivated Lyn following Ag stimulation, resulting in the suppressions of intracellular elevation of [Ca²⁺]i and production of ROS. To identify the active compound in EtEx.40, we isolated 7 flavonoids from EtEx.40 and calculated their inhibition levels on Ag-stimulated degranulation. These flavonoids inhibited degranulation by about 25-60%. We further examined the in vivo effects of HWEA or EtEx.40 using a passive cutaneous anaphylaxis (PCA) reaction. Both extracts strongly suppressed the PCA reaction. These findings suggest that HWEA and/or EtEx.40 are beneficial for alleviating type I allergic symptoms.

The absolute configration of the epoxy group in 13β,17β:24(R),25-diepoxy-1β-hydroxydammar-3-one 23(S)-acetate, C 32 H 50 O 6 , m.p. 472-474 K, [α] D +139.4 o (c=0.96, CHCl 3 ), isolated from Alisma orientale rhizomes, was established to be the β orientation by X-ray crystallographic analysis. The A, B and C rings have chair forms. An intermolecular hydrogen bond is observed between O2 and O6' with a distance of 3.148 (6) A

The structure of a new diterpene, C 20 H 30 O 2 , m.p. 452-453 K, [α] D -28.4 o (c=1.0, acetone), isolated from fresh rhizomes of Alisma orientale (Alismataceae) was established to be 16(R)-(-)-kaurane-2,12-dione by means of X-ray crystallographic analysis and its optical rotation. The A, B and C rings have chair conformations. The puckering parameter of the bridging D ring, the angle between the planes C(14)-C(13)-C(16)-C(15) and C(15)-C(8)-C(14), is 39.4 (7) o

Two new saponins, achyranthosides A and B, were isolated from Achyranthes fauriei root, and their structures were elucidated on the basis of chemical and physical evidences. Achyranthoside A methyl ester was found to have significant cytotoxic activity against human colon carcinoma and murine melanoma cells.

The structures of Julibrosides I, II, and III, isolated from dried stem bark of Albizzia julibrissin DURAZZ (Albizziae Cortex), were characterized as complex triterpenoidal glycosides linked to the monoterpene glycoside dimer, mainly by means of an NMR spectroscopic method. These compounds have a common structure comprising from acacic acid, the (6S)-2-trans-6-hydroxy-2,6-dimethy1-2,7-octadienoyl-6-O-quinovopyranoside dimer, and α-L-arabinofuranosyl-(1→4)-[β-D-glucopyranosyl-(1→3)]-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside. The diversity of the oligosaccharides attached to the C-3 hydroxyl group of acacic acid, an oleanane-type triterpene, makes a distinction between Julibrosides I, II, and III.