Synthesis of dihydroxyphenacyl glycosides for biological and medicinal study: β-oxoacteoside from Paulownia tomentosa

The protected structure of -oxoacteoside (tomentoside A), 2-oxo-2-(3,4-dihydroxyphenyl)ethyl 3-O-(2,3,4-tri-O-acetyl--l-rhamnopyranosyl)-4-O-caffeoyl--d-glucopyranoside 14 was synthesized in 14% overall yield in 11 steps, starting from d-glucose for biological and medicinal studies of phenylpropanoid glycosides. The first step was the preparation of a 3-O-rhamnopyranosyl disaccharide sugar core 2 from a suitably protected rhamnosyl trichloroacetimidate 10 and glucose derivative (diacetone-d-glucose 1) in 71% yield. To the glucose moiety of this sugar core, several protection/deprotection procedures were performed sequentially to obtain a fully acetylated sugar core 7 with a 4-OH group on the glucose moiety, in 57% yield in five steps. Thereafter, to the 4-OH group of the glucose moiety, selective 4-O-caffeoylation was achieved by proton-transfer esterification with 3,4-di-O-allylcaffeic acid 16 to give the caffeoyl disaccharide 11 in 97% yield. Then, it was converted to trichloroacetimidate 13 for a glycosylation donor in 90% in two steps. Finally, anomeric glycosylation was conducted with 2-oxo-2-(3,4-di-allyloxyphenyl)ethyl alcohol 19 with catalytic amounts of BF₃·Et₂O to give 2-oxo-2-(3,4-di-allyloxyphenyl)ethyl 2,6-di-O-acetyl-3-O-(2,3,4-tri-O-acetyl--l-rhamnopyranosyl)-4-O-(3,4-di-allyloxycaffeoyl)--d-glucopyranoside 14 in 60% yield. Deprotected intermediates of compounds 2, 11, 14, and 19 which were obtained in high yield would be useful for biological and medicinal studies of phenylpropanoid glycosides.Part of this study was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April, 2002     

Antioxidative compounds from leaves of Tahongai (<Emphasis Type="Italic">Klienhovia hospita</Emphasis>)

In our effort to find antioxidant agent, we focused on Tahongai (Kleinhovia hospita) which have been used traditionally in Indonesia as medicinal herbal to cure liver disease. Based on the biologically guided fractionation using DPPH radical scavenging assay, eleutherol and kaempferol 3-O-β-d-glucoside was isolated from the leaves of K. hospita. Kaempferol 3-O-β-d-glucoside (1) and eleutherol (2) scavenged the radical with IC₅₀ of 71.4 and 491.8 μM, respectively. In addition, both of the compounds did not exhibit cytotoxicity on HepG2 cells.     

Chemical constituents from Gmelina arborea bark and their antioxidant activity

Gmelina arborea Roxb. is a fast-growing species and is known to have been used in traditional Indian medicine. Chemical constituents from the bark have not been reported, although some chemical constituents from part of this plant (heartwood, leaf, and root) are known. In this study, the bark meal was successively extracted with acetone and methanol. Fractionation of the acetone extract with n-hexane, diethyl ether, and ethyl acetate and subsequent chromatographic separation of the fractions led to the isolation of four compounds. The diethyl ether-soluble fraction yielded tyrosol [2-(4-hydroxyphenyl)ethanol] (1); (+)-balanophonin (2), an 8-5′ neolignan, with opposite optical rotation to known (−)-balanophonin; and gmelinol (3), a known lignan. The ethyl acetate-soluble fraction afforded a new phenylethanoid glycoside to the best of our knowledge, which was identified as (−)-p-hydroxyphenylethyl[5′″-O-(3,4-dimethoxycinnamoyl)-β-d-apiofuranosyl(1′" → 6′)]-β-d-glucopyranoside (4). From the methanol extract, two known compounds, 2,6-dimethoxy-p-benzoquinone (5) and 3,4,5-trimethoxyphenol (6), were isolated and identified. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay of the identifi ed compounds indicated that 3,4,5-trimethoxyphenol (6) exhibited moderate activity. Part of this report was presented at the 57th and 58th Annual Meetings of the Japan Wood Research Society, Hiroshima and Tsukuba, August 2007 and March 2008, respectively     

Structural unit of xylans from sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) and hinoki (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>)

Arabinoglucuronoxylans (AGXs) isolated from the holocellulose of sugi (Cryptomeria japonica) and hinoki (Chamaecyparis obtusa) contained one 4-O-methyl-d-glucopyranosyluronic acid (4-O-Me-d-GlcAp) residue per 6.2 d-xylopyranose (d-Xylp) residues and one 4-O-Me-d-GlcAp residue per 3.8 d-Xylp residues. These AGXs were subjected to partial acid hydrolysis. Analyses by size exclusion chromatography and electrospray-ionization mass spectroscopy of the neutral sugar fractions in the hydrolysates showed the presence of xylooligosaccharides having a degree of polymerization of 2-8 in addition to d-Xyl, suggesting that the AGXs from sugi and hinoki contained unsubstituted chains consisting of at least eight d-Xyl residues. The acidic sugars in the hydrolysates were separated into two series of aldouronic acids composed of 4-O-Me-d-GlcAp and d-Xylp by ion-exchange chromatography. The first series included aldouronic acids from aldobiouronic acid (4-O-Me-d-GlcAp-Xyl) to aldopentaouronic acids (4-O-Me-d-GlcAp-Xyl₄). The second series were aldouronic acids composed of two 4-O-Me-d-GlcAp residues and 2-4 d-Xyl residues. In these acidic sugars, the uronic acid side chains were located on two contiguous d-Xyl residues. These facts indicated that AGXs from sugi and hinoki had a structural unit containing two 4-O-Me-d-GlcAp residues on two contiguous d-Xyl residues as well as AGXs from spruce and larch.     

Investigation on hydrogen abstraction from methyl glucoside by active oxygen species under oxygen delignification conditions III: effects of the origin of active oxygen species

Carbohydrate model compounds methyl β-d-glucopyranoside (MGPβ), methyl α-d-glucopyranoside (MGPα), and methyl β-d-mannopyranoside (MMPβ) and the deuterium compounds of MGPβ labeled at the anomeric or C-2 positions (MGPβ-1D, MGPβ-2D) were reacted with active oxygen species (AOS) generated in situ by reactions between O₂ and a co-treated phenolic lignin model compound, 4-hydroxy-3-methoxybenzyl alcohol (VAlc), under conditions simulating oxygen delignification (0.5 mol/l NaOH, 0.36 mmol/l Fe³⁺, 1.1 MPa O₂, 95°C). MGPβ was degraded more than MGPα but less than MMPβ when the pairs MGPβ/MGPα and MGPβ/MMPβ, respectively, were treated, which indicates that the configurational differences at the anomeric and C-2 positions influence the reactivity of AOS toward these compounds. When the pairs MGPβ/MGPβ-1D and MGPβ/MGPβ-2D were treated, no clear kinetic isotope effects were observed in either case. These results contrasted with those obtained when another phenolic compound, 2,4,6-trimethylphenol (TMPh), was used as the AOS generator instead of VAlc under exactly the same conditions. Clear kinetic isotope effects were observed when using TMPh. Because it is not easily accepted that the anomeric and C-2 hydrogen abstractions are minor reaction modes only for AOS generated in the VAlc system, it is suspected that the AOS do not show any clear kinetic isotope effect even though the AOS abstract an objective hydrogen.     

Antioxidant properties and structural analysis of phenolic glucosides from bark of <Emphasis Type="Italic">Populus ussuriensis</Emphasis> Kom.

The methanol (MeOH) extract of Populus ussuriensis Kom. bark was analyzed for antioxidant assessing by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging potential. Among fractions using several solvents, the ethyl acetate (EtOAc) soluble fraction, which showed strong antioxidant activity (IC₅₀ 2.02 ± 0.01 μg/ml), was further purified by Thin layer chromatography (TLC) guided Sephadex LH-20 column chromatography. Three known phenolic glucosides, picein (I), salicortin (II), grandidentatin (III), and that of a new, 2-hydroxycyclohexyl-4′-O-p-coumaroyl-β-d-glucopyranoside (isograndidentatin A), were isolated and their structures were elucidated on the basis of physiochemical and spectroscopic methods. This was the first report of the chemical composition of P. ussuriensis bark. Phenolic glucosides III and IV exhibited strong antioxidant activities, with IC₅₀ values 6.73 ± 0.01 and 6.69 ± 0.01 μM, respectively, comparable to the control (α-tocopherol, IC₅₀ 6.80 ± 0.01 μM). P. ussuriensis bark EtOAc-soluble fraction and compounds III and IV could be used as biopreservatives in food applications as well as for cosmetic and medicinal preparations, to alleviate oxidative stress.     

Four glucosides ofp-hydroxyphenyl derivatives from birch leaves

The extractive of shirakamba (Betula platyphylla Sukatchev var.japonica Hara) leaves was investigated. Four glucosides ofp-hydroxyphenyl derivatives were isolated, and their structures were indentified as betuloside (I), 3,4-dihydroxy-propiophenone-3--d-glucopyranoside (II), salidroside (III), and arbutin (IV). Arbutin was newly found in the leaves of shirakamba.     

A glucosylceramide with antimicrobial activity from the edible mushroom <Emphasis Type="Italic">Pleurotus citrinopileatus</Emphasis>

The golden oyster mushroom Pleurotus citrinopileatus is a popular edible mushroom with multifunctional biological activities, but there are a limited number of previous studies on its chemical composition. This is the first report of the isolation of glucosylceramide with antimicrobial activity from the fruiting body of this mushroom. This compound was identified as 1-O-β-d-glucopyranosyl-(2S,3R,4E,8E)-2-[(2R)-2-hydroxyhexadecanoylamino]-9-methyl-4,8-octadecadiene-1,3-diol. The IC₅₀ value of this compound for the growth of Escherichia coli and Staphylococcus aureus was 275.1 μM (200 μg/ml) and 323.2 μM (235 μg/ml), respectively.     

Effects of stilbenes from bark of <Emphasis Type="Italic">Picea glehnii</Emphasis> (Sieb. et Zucc) and their related compounds against feeding behaviour of <Emphasis Type="Italic">Reticulitermes speratus</Emphasis> (Kolbe)

The effects of stilbene glucosides and related compounds on termite feeding behavior were investigated using paper disc methods against the subterranean termite Reticulitermes speratus. The stilbene-rich fraction and isorhapontin (3-methoxy-3,4, 5-trihydroxystilbene-3--d-glucoside) from bark extracts of Picea glehnii showed avoidance by termites in choice tests. In the no-choice tests using compounds purified from the stilbene-rich fraction, the largest feeding deterrent effect was observed for piceid (3,4,5-trihydroxystilbene-3--d-glucoside), followed by isorhapontin, and astringin (3,3,4,5-tetrahydroxystilbene-3--d-glucoside), at the concentrations from 0.63 to 2.5µmol/disc. No change in activity was observed at retentions of more than 5.0µmol/disc. When the activities of isorhapontin and its aglycone derivative (isorhapontigenin: 3-methoxy-3,4,5-trihydroxystilbene) were compared with that of taxifolin (3,3,4,5,7-pentahydroxyflavanone) in the no-choice test, the stilbenes exhibited a larger antifeedant potential. Methylation of isorhapontigenin increased its termiticidal activity.Part of this study was presented at the 32nd Annual Meeting of the International Research Group on Wood Preservation, 2001     

Role of caffeic glucoside esters in defense-repair processing of trees II: Synthesis of 2-(3,4-dihydroxyphenyl)-ethyl 3-α-L-rhamnopyranosylβ-D-glucopyranoside

The reference compound (9), with a partial structure of acteoside, was synthesized to elucidate the relation between structural features and the precipitation or solubility of the oxidation products of acteoside: 2-(3,4-dihydroxyphenyl)-ethyl 3-O-(-L-rhamnopyranosyl)--D-glucopyranoside (9). The glycosyl acceptor 2-O-benzoyl-3-O-(2,3,4-tri-O-acetyl--L-rhamnopyranosyl)-4,6-O-benzylidene--D-glucopyranosyl trichloroacetimidate (7) was prepared from allyl 2-O-benzoyl-3-O-(2,3,4-tri-O-acetyl--L-rhamnopyranosyl)-4,6-O-benzylidene--D-glucopyranoside (4) via isomerization of the allyl group with an iridium complex to the 1-propenyl group and its hydrolysis with HgCl₂lHgO, followed by treatment with CCl₃CN and DBU in a 65.5% overall yield. The glycosyl acceptor 3,4-diacetoxyphenethyl alcohol (16) was prepared from homoveratric acid via demethylation with 57% HI and its acetylation with Ac₂O and 85% H₃PO₄, followed by selective reduction of the carboxyl group to the alcohol with a borane-tetrahydrofurane complex in a 61% overall yield. The glycosylation of7 with16 in dichloromethane promoted by BF₃-Et₂O gave a 74.3% yield of8. Hydrolysis of8 with 90% CF₃COOH gave the debenzylidenated product, which was treated with NaOMe to afford a 32% overall yield of the desired compound9. This compound9 was identical with the natural specimen.This study was presented in part at the 46th annual meeting of the Japan Wood Research Society, Kumamoto, April 3–5, 1996     

Microwave pyrolysis of cellulosic materials for the production of anhydrosugars

Large-scale microwave rapid pyrolysis of cellulosic materials has been investigated. Levoglucosan (1,6-anhydro--d-glucopyranose) was obtained from a larch log as the main anhydrosugar in 2.6% yield on the basis of dry wood weight. This yield would be much higher than that obtainable by conventional pyrolysis in the largescale reaction. Levoglucosenone (1,6-anhydro-3,4-dideoxy--D-glycero-hex-3-enopyranos-2-ulose) was found to be produced in one-quarter the amount of levoglucosan. Other anhydrosugars, such as mannosan (1,6-anhydro--D-mannopyranose), galactosan (1,6-anhydro--d-galactopyranose), and xylosan (1,4-anhydro--d-xylopyranose), were also confirmed to be produced as minor components depending on the proportion of the monosaccharide content in the larch. When microwave pyrolysis of used papers and filter papers was performed, the yields of levoglucosan were about 6% and 12%, respectively, suggesting that a higher content of cellulose gives a larger amount of levoglucosan.     

Antihyperglycemic effects of Japanese maple Acer amoenum leaf extract and its constituent corilagin

The antihyperglycemic effects of the leaves of Acer amoenum and purification and identification of an active compound were investigated. In screening experiments for α-glucosidase inhibitory activity, methanolic extracts of A. amoenum leaves showed potent inhibitory action. This extract showed antihyperglycemic effects in sucrose-loaded mice. Fractionation of the crude extract gave the active compound corilagin [β-1-O-galloyl-3,6-(R)-hexahydroxydiphenoyl-d-glucose] by spectroscopic analysis. This is the first report about the possibility of novel utilization of the Japanese maple tree as a source of compounds for prevention or treatment of diabetes mellitus.     

Synthesis of isoacteoside, a dihydroxyphenylethyl glycoside

The total chemical synthesis of isoacteoside (1), 2-(3,4-dihydroxyphenyl)ethyl 6-O-caffeoyl-3-O-(-l-rhamnopyranosyl)--d-glucopyranoside, is described. An acteoside acetate with benzyl groups at the catechols (3: 2-(3,4-dibenzyloxyphenyl)ethyl 2,6-di-O-acetyl-4-O-[3,4-bis(O-benzyl)caffeoyl]-3-O-(-l-rhamnopyranosyl)--d-glucopyranoside) was treated with a solution of methy-lamine in methanol (MeNH₂ in MeOH) to perform both deacetylation and caffeoyl migration, affording an isoacteoside derivative with benzyl groups at the catechols4b: 2-(3,4-dibenzyloxyphenyl)ethyl 6-O-[3,4-bis(O-benzyl) caffeoyl] -3-O-(-l-rhamnopyranosyl)--d-glucopyranoside —in 34% yield. Debenzylation of4b was successfully accomplished by catalytic transfer hydrogenation using 1,4-cyclohexadiene to give the target compound isoacteoside (1) in 54% yield.¹H and¹³C nuclear magnetic resonance spectral data of the synthesized isoacteoside (1) were identical with those of the natural isoacteoside isolated fromPaulownia tomentosa (Thumb.) Steud.Part of this research was presented at the 51st Annual Meeting of the Japan Wood Research Society, Tokyo, April 2001     

Influences of nitrogen and phosphorus addition on polyphenol oxidase activity in a forested Andisol

Increased atmospheric N deposition could suppress plant litter decomposition, due to the P limitation for soil microorganisms in Japanese forested Andisols with a high P sorption capacity. To explore this possibility, we used a laboratory incubation experiment to study the influence of N addition on β-d-glucosidase and polyphenol oxidase activities, which are important for cellulose and lignin degradation, respectively, in an Andisol with larch (Larix kaempferi) leaf litter. The addition of N increased the β-d-glucosidase activity, whereas it decreased the polyphenol oxidase activity in the soil. However, the addition of both N and P increased the polyphenol oxidase activity in the soil, suggesting the possibility of; (1) an inferior competitive ability of polyphenol oxidase-producing microorganisms under nutrient-rich conditions and; of (2) their P limitation through competition in the Andisol.     

Structure of xylan from culms of bamboo grass (Sasa senanensis Rehd.)

Xylan prepared from culms of kumaizasa (Sasa senanensis Rehd.), a representative species of bamboo grass, was hydrolyzed with-xylanase ofStreptomyces olivaceoviridis E-86. Four arabinoxylo-oligosaccharides and two glucuronoxylo-oligosaccharides were isolated from the enzymatic hydrolysate of the xylan by chromatography on a charcoal column, a Dowex 1-x8 column, a Toyo-pearl HW-40S column, and a LiChrospher 100 NH₂ column and on preparative paper chromatography. The results of the structural analyses of the saccharides showed that the isolated oligosaccharides had the structures of 3²--l-arabinofuranosyl-xylobiose, 3²--l-arabinofuranosyl-xylotriose, 3²--[-d-xylopyranosyl-(1 2)-l-arabinofuranosyl]-xylobiose, 3³--[-d-xylopyranosyl-(1 2)-l-arabinofuranosyl]-xylotriose, 2³--4-O-methyl-d-glucuronosyl-xylotriose, and 2³--d-glucuronosyl-xylotriose. From the structural analysis of the oligosaccharides derived from the xylan, kumaizasa xylan was concluded to be a kind of arabinoglucuronoxylan having not only stubs of singlel-arabinose and singled-glucuronic acid but also stubs of disaccharide units such as-d-xylopyranosyl-(1 2)-l-arabinofuranose.     

Sulfated glycofuranans as inhibitors of melanoma lung metastasis

Chemically synthesized (1 5)--d-glucofuranan, (1 5)--d-galactofuranan, (1 5)--d-xylofuranan, (1 5)--L-arabinofuranan, natural xylan, and curdlan were sulfated to investigate their inhibitory activities on B16-BL6 lung metastasis and anticoagulant activities. (1 5)--d-Glucofuranan sulfate, (1 5)--d-galactofuranan sulfate, xylan sulfate, and curdlan sulfate had binding abilities with B16-BL6 melanoma lysate. The inhibitory activities of sulfated polysaccharides on B16-BL6 lung metastasis selected by heparin binding assay were in the order (1 5)--d-galactofuranan sulfate > (1 5)---d-glucofuranan sulfate > xylan sulfate curdlan sulfate. Furthermore, (1 5)--d-galactofuranan sulfate, (1 5)--d-glucofuranan sulfate, and xylan sulfate had not only high inhibitory activity on B16-BL6 lung metastasis but also low anticoagulant activity. The correlation between chemical structure and biological activity is discussed.Part of this paper was presented at the 10th International Synposium on Wood and Pulping Chemistry, Yokohama, Japan, June 1999     

A new iridoid from Adenosma caeruleum R. Br.

A new iridoid glycoside, adenosmoside (1), together with five known phenylpropanoids, crenatoside, verbascoside, cistanoside F, campneoside I, and campneoside II and two known flavonoids, apigenin 7-O-β-d-glucuronopyranoside and apigenin 7-O-β-d-glucopyranoside, were isolated from the aerial parts of Adenosma caeruleum R. Br. Their structures were elucidated by spectral evidence.     

Preparation of novel reagents 4-alkoxytrityl chlorides and their reaction with methyl Α-d-glucoside

A series of novel 4-O-alkoxytrityl chlorides (1) with different chain lengths was synthesized as a novel reagent for obtaining 6-O-alkylated cellulose with high regioselectivity via trityl groups in one reaction step without the use of any protective groups. These chlorides were reacted with methyl -d-glucoside, which was used as a model compound, to examine the reactivities toward the primary hydroxyl groups of cellulose to afford a series of 6-O-alkylated methyl -d-glucosides in high yields. The product compounds were found to have interesting solubilities and thermal properties. Thus, newly prepared trityl chloride derivatives were found to be useful regioselective derivatization reagents on the primary hydroxyl group in carbohydrates, especially in cellulose.     

Isolation of a β-1,3-glucan (laricinan) from compression wood of Larix laricina

Summary A -d-(13)-linked glucan has been isolated from compression wood of tamarack [Larix laricina (Du Roi) K. Koch]. The polysaccharide, which had []_D + 17.6°, consumed 0.17 mole of periodate per glucose residue and gave a methyl derivative which consisted of 115 2,4,6-tri- and 2,3,4,6-tetra-O-methyl-d-glucose residues in a ratio of 17.8:1. Partial acid hydrolysis yielded a series of -d-(13)-linked oligosaccharides, while enzymic hydrolysis with a -1,3-glucanase afforded glucose, laminaribiose, and laminaritriose. The glucan occurred in amounts varying from 2 to 4% in compression wood of this and other conifer species and was present in the wall of both tracheids and ray cells. It is proposed that this structural polysaccharide be designated as laricinan to distinguish it from callose and the various -1,3-glucans in lower plants.     

Neuropharmacological activities of phytoncide released from <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

Forest-air bathing and walking (shinrin-yoku) is beneficial to human heath. In this study the phytoncide (volatile compounds) released from Cryptomeria japonica plantation forest was characterized by using solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). The main volatile compounds were α-pinene (19.35%), β-myrcene (16.98%), d-limonene (15.21%), and γ-muurolene (7.42%). Furthermore, the neuropharmacological activity of the essential oils from leaves of C. japonica (ECJ) was evaluated by several animal behavior tests. ECJ could prolong the sleeping phase of ICR (imprinting control region) mice in the pentobarbital-induced sleeping time model. Furthermore, both EJC and one of its monoterpenes, d-limonene, possessed potent anxiolytic and analgesic activities based on the results obtained from elevated plus maze and writhing tests. The volatile compounds released from C. japonica provide relaxing and stress-relieving effects on mice, and further study on the effect of phytoncide on humans is worthwhile.