Crataegusins A and B, New Flavanocoumarins from the Dried Fruits of Crataegus pinnatifida var. major (Rosaceae).

Crataegusins A (1) and B (2), new flavanocoumarins, were isolated from the crude drug Crataegus Frictus, i.e., the dried fruits of Crataegus pinnatifida var. major..Their structures were determined by spectroscopic methods. They were unique in terms of carrying a 3-(or 4-)substituted coumarin substructure while a flavanocoumarin generally does not carry any substituents in the 2-pyron ring. They showed a significant DPPH reducing activity compared with epicatechin Their production would be biosynthetically regulated considering the results of an LC-MS analysis of the dried and fresh fruits, fruit skin, hypanthia, and leaves. Their structures led the authors to consider a hypothetical general biosynthetic pathway of the flavanocoumarins, to which a flavan-3-ol is converted through a Michael addition and successive oxidative decarboxylation or dehydration pathway.

Novel Monoterpene Lactones from Cinnamomum inunctum.

Four new monoterpene lactones, 5-(2,3-dihydroxy-3-methylbutyl)-4-hydroxy-4-methyldihydrofuran-2(3H)-one (1), 5-(2,3-dihydroxy-3-methylbutyl)-4-methylfuran-2(5H)-one (2), 8-hydroxy-4,7,7-trimethyl-1,6-dioxaspiro[4.4]non-3-en-2-one (3) and 8-hydroxy-4,7,7-trimethyl-1,6-dioxaspiro[4.4]non-3-en-2-one (4), were isolated from the methanolic extract of the fruit of Cinnamomum inunctum, a folk medicine in Myanmar, together with a known compound, 3-hydroxy-4,4-dimethyl-4-butyrolactone (5). Their chemical structures were determined by spectral methods. Among these, 3 and 4 possessed unique spirolactone moieties.

New leishmanicidal stilbenes from a Peruvian folk medicine, Lonchocarpus nicou.

Two new stilbenes, 1 and 2, were isolated as leishmanicidal constituents from the methanolic extract of Lonchocarpus nicou leaves and stem, together with five known stilbenes and rotenoids. Their chemical structures were determined by spectral methods. Among them, the cis stilbene-type compounds 1 and 4 showed potent leishmanicidal activity (IC50: 5.5, 3.9 µg/mL), while the trans stilbene-type compounds 2 and 5, and rotenoids 6 and 7, showed moderate activities (IC50: 9.9, 12.8, 22.6, 19.6 µg/mL, respectively).

Leishmanicidal activities and cytotoxicities of bisnaphthoquinone analogues and naphthol derivatives from Burman Diospyros burmanica.

A methanol extract of the wood of Diospyros burmanica, collected in Burma (Myanmar), was found to exhibit significant activity against Leishmania major. Subsequent chromatographically resolved fractionation led to the isolation of three novel bisnaphthoquinone analogues, burmanin A, B, and C (1-3), together with nine known compounds (4-12). The structure of 1 was confirmed by X-ray crystallography, and those of 2 and 3 by spectroscopic techniques, including 1D and 2D NMR. The inhibitory activities of the isolates were evaluated against the promastigote forms of Leishmania major and the murine macrophage-like cell line, RAW264.7.

Sesquiterpenes from Blumea balsamifera.

Five new guaiane sesquiterpenes, blumeaenes E1 (1), E2 (2), K (3), L (4), and M (5), and one new eudesmane sesquiterpene, samboginone (6), along with three known compounds, cryptomeridiol, 3,3',5,7-tetrahydroxy-4'-methoxyflavanone, and austroinulin, were isolated from the leaves of the Philippine medicinal herb sambong, Blumea balsamifera. The absolute configuration of the new guaiane core was determined as 1S,7S,9S,10R by employing the modified Mosher's method. In the structure of 1, the absolute configuration of the epoxyangelic acid moiety was identified as 2S,3S using (R)-PGME as a chiral anisotropic auxiliary.

In vitro leishmanicidal activity of benzophenanthridine alkaloids from Bocconia pearcei and related compounds.

Leishmanicidal activities of benzophenanthridine alkaloids isolated from fruits of Bocconia pearcei and their derivatives were examined. Seven benzophenanthridine compounds were isolated from the methanolic extracts of B. pearcei. Among them, dihydrosanguinarine showed the most potent leishmanicidal activities (IC(50) value: 0.014 microg/ml, respectively). To examine the structure-activity relationship of the benzophenanthridine skeleton, the leishmanicidal activities for 32 synthetic samples were examined. The existence of bulky groups at the C(7)-C(8) position was found to enhance the activity. On the other hand, the bulkiness at the C(2)-C(3) position on the D-ring, a carbonyl group at C-6, substitution at C-6 and cleavage or saturation of the C(5)-C(6) bond reduced activity. A methyl group on nitrogen of the C-ring was thought to be necessary for significant activity.

Identification of medicinal Dendrobium species by phylogenetic analyses using matK and rbcL sequences.

Species identification of five Dendrobium plants was conducted using phylogenetic analysis and the validity of the method was verified. Some Dendrobium plants (Orchidaceae) have been used as herbal medicines but the difficulty in identifying their botanical origin by traditional methods prevented their full modern utilization. Based on the emerging field of molecular systematics as a powerful classification tool, a phylogenetic analysis was conducted using sequences of two plastid genes, the maturase-coding gene (matK) and the large subunit of ribulose 1,5-bisphosphate carboxylase-coding gene (rbcL), as DNA barcodes for species identification of Dendrobium plants. We investigated five medicinal Dendrobium species, Dendrobium fimbriatum, D. moniliforme, D. nobile, D. pulchellum, and D. tosaense. The phylogenetic trees constructed from matK data successfully distinguished each species from each other. On the other hand, rbcL, as a single-locus barcode, offered less species discriminating power than matK, possibly due to its being present with little variation. When results using matK sequences of D. officinale that was deposited in the DNA database were combined, D. officinale and D. tosaense showed a close genetic relationship, which brought us closer to resolving the question of their taxonomic identity. Identification of the plant source as well as the uniformity of the chemical components is critical for the quality control of herbal medicines and it is important that the processed materials be validated. The methods presented here could be applied to the analysis of processed Dendrobium plants and be a promising tool for the identification of botanical origins of crude drugs.

Simple and rapid analysis of aristolochic acid contained in crude drugs and Kampo formulations with solid-phase extraction and HPLC photodiode-array detection.

Simple and rapid analysis of aristolochic acid (AA) in crude drugs and Kampo extracts using a solid-phase extraction method and HPLC-PDA analysis was investigated. Extraction of AA from samples was accomplished by adding methanol containing 1% ammonia. The addition of ammonia ionized the AA of acidic substances so that they adhered to an acrylamide copolymer of a strong anion exchange resin (Sep-Pak QMA) coupled to diol silica easily. Furthermore, a mixture of acetonitrile-water-phosphoric acid (75:25:2, v/v) was effective in isolating AA from its carrier. Since almost all interfering peaks originating from contaminants in crude drugs and Kampo extract formulations could be removed, a satisfactory HPLC chromatogram of AA was obtained. A good result was also obtained when Aristolochiaceae and crude drugs containing AA were tested. Particularly in the case of the medicinal parts of Asarum, several interfering peaks and a ghost peak detected near the AA peak were eliminated. The AA contents of two Kampo extract formulations, tokishigyakukagoshuyushokyoto and ryutanshakanto, were calculated by HPLC analysis. The AA content (the sum of AA-I and AA-II) was 1.25-6.13 mg per daily dose. From an additional recovery experiment for Kampo formulations, high recovery rates of AA were obtained. Neither LC/MS nor special instrumentation was necessary. Our results suggest that this simple, quick, and sensitive analytical method to detect AA in crude drugs and Kampo extract formulations would be valuable in safety inspections of AA in crude drugs and their products.

Anti-inflammatory and analgesic components from "hierba santa," a traditional medicine in Peru.

"Hierba santa," a Peruvian herbal medicine, is used to alleviate many symptoms, including headache, hemorrhoids, fever, and rheumatism. Several Cestrum species are said to be the origin of hierba santa. Three lots of hierba santa: Cestrum auriculatum (herb 1 and herb 2) and C. hediundinum (herb 3), which were purchased from Peruvian markets at Cuzco (Andes area) and Equitos (Amazon area), respectively, were examined for their pharmacological activities and active components. Herbs 1-3 showed anti-inflammatory and analgesic activities in the in vivo writhing inhibition test in mouse and inhibited prostaglandin E(1)-, E(2)-, or ACh-induced contractions of guinea pig ileum in the Magnus method. Activity-based separation of each extract yielded cestrumines A and B, cestrusides A and B, a mixture of (+)- and (-)-pinoresinol glucosides, nicotiflorin, rutin, sinapoyl glucose, ursolic acid, beta-sitosteryl glucoside, and 2-sec-butyl-4,6-dihydroxyphenyl-beta-D: -glucopyranoside. Among them, cestrumine A and cestrusides A and B are new compounds. All three lots of hierba santa do not contain exactly the same active components.

A new leishmanicidal saponin from Brunfelsia grandiflora.

A new furostan-type saponin (1) was isolated from the methanolic extract of Brunfelsia grandiflora leaves, together with four known compounds. The chemical structure of 1 was determined by spectroscopic analysis and chemical reaction to be 26-O-beta-D-glucopyranosyl 22alpha-methoxyfurost-3beta,26-diol 3-O-beta-D-xylopyranosyl(1-->3)-{beta-D-glucopyranosyl(1-->2)}-beta-D-glucopyranosyl(1-->4)-beta-D-glucopyranoside. Compound 1 showed potent leishmanicidal activity in vitro against Leishmania major.

Antileishmanial compounds from Cordia fragrantissima collected in Burma (Myanmar).

A methanol extract of the wood of Cordia fragrantissima, collected in Burma (Myanmar), was found to exhibit significant activity against Leishmania major. Bioassay-guided fractionation of this extract using several chromatographic techniques afforded three new compounds (1-3) and five known compounds (4-8). The structures of the new compounds were revealed on the basis of spectroscopic data interpretation and by X-ray crystallographic analysis. Interestingly, the new compounds, despite the presence of asymmetric carbons, were found to be racemates. The activities of the isolates from C. fragrantissima and several derivatives were evaluated against the promastigote forms of Leishmania major, L. panamensis, and L. guyanensis.

In vitro leishmanicidal constituents of Millettia pendula.

The in vitro leishmanicidal constituents of Millettia pendula were examined. Two new compounds, 1 (millettilone A) and 2 (millettilone B), were isolated from the methanol extract of M. pendula, together with six known compounds: 3R-claussequinone (3), pendulone (4), secundiflorol I (5), 3,8-dihydroxy-9-methoxypterocarpan (6), 3,10-dihydroxy-7,9-dimethoxypterocarpan (7), and formononetin (8). Among these, pendulone showed the most potent leishmanicidal activity. Compound 2 was found to be a purple pigment in this heartwood. Their chemical structures were elucidated using spectral methods.

Two cangorosin A type triterpene dimers from Maytenus chuchuhuasca.

Two new cangorosin A type triterpene dimers, which composed of two triterpene units jointed by two ether linkages between the A and B rings, were isolated from the Brazilian medicinal plant "xuxuá" (Maytenus chuchuhuasca). Structures of new isolates, xuxuasins A (1) and B (2), were established based on several spectroscopic evidences.

Two new cucurbitacin glucosides, opercurins A and B, from the Brazilian folk medicine "Buchinha" (Luffa operculata).

The structures of two new cucurbitacin glucosides designated opercurins A (1) and B (2), isolated from the fruit of Luffa operculata, have been confirmed by extensive spectroscopic investigation.

In vitro leishmanicidal activity of some scarce natural products.

Leishmanicidal activity of 46 natural products including several fern and Betula constituents was examined. Several pterosin and atisene compounds from ferns had high activity. Among the triterpenoids the carboxyl group was found to be important for activity. In the diarylheptanoids, the linear-type and the diphenylether-type showed significant activity, and it was found that the carbonyl group at C-11 is necessary for the activity of biphenyl-types.

Nine regioisomeric and stereoisomeric triterpene dimers from Maytenus chuchuhuasca.

Nine regioisomeric and stereoisomeric triterpene dimers, namely xuxuarine Falpha (1), isoxuxuarine Falpha (2; cangorosin B), 7,8-dihydroisoxuxuarine Falpha (3), isoxuxuarine Gbeta (4), 7,8-dihydroisoxuxuarine Galpha (5), isoxuxuarine Ebeta (6), 7alpha-hydroxyisoxuxuarine Ealpha (7), 7',8'-dihydroxuxuarine Aalpha (8), and 7',8'-dihydroxuxuarine Dbeta (9), were isolated from the Brazilian medicinal plant "xuxuá" (Maytenus chuchuhuasca). Their structures have been elucidated based on several spectroscopic analyses including 2D-NMR experiments, MS spectra and CD spectral studies.

In vitro screening of leishmanicidal activity in myanmar timber extracts.

Seventy-five Myanmar timber extracts belonging to 27 families were examined for their leishmanicidal activities. Some timber extracts had significant leishmanicidal activity, especially extracts of Millettia pendula, which exhibited the most potent activity (MLC 3.1 microg/ml, MIC 1.6 microg/ml). Other timber extracts showing potent activity included those from Cedrela serrata, Cedrela toona, Cordia fragrantissima, Calophyllum kunstleri, Dalbergia cultrate, Grevillea robusta, Haplophragma adenophyllum, Michelia champaca, and Tectona grandis. From a literature search for reports on the chemical constituents of these plants, most constituents were found to be quinone derivatives or other compounds with unsaturated carbonyl groups.

Nine new isoxuxuarine-type triterpene dimers from Maytenus chuchuhuasca.

Nine new isoxuxuarine-type triterpene dimers, named isoxuxuarines B alpha (1), B beta (2), 7,8-dihydro-B alpha (3), C alpha (4), C beta (5), 7,8-dihydro-C alpha (6), D alpha (7), D beta (8), and 7,8-dihydro-D alpha (9), were isolated from the South American medicinal plant 'xuxuá' (Maytenus chuchuhuasca). The structures were elucidated on the basis of several spectroscopic analyses, including 2D-NMR experiments, MS spectra, and CD spectral studies. The isolations and structure elucidations of the new triterpene dimers are presented, and a rationale for the divergent formation of the regiochemical and stereochemical isomers of triterpene dimers is discussed.

Dammarane-type triterpenes from the Brazilian medicinal plant Cordia multispicata.

From the Brazilian medicinal plant Carucaá (Cordia multispicata), oleanane- and ursane-type triterpenoids were previously reported as anti-androgenic constituents of the plant. In this study, purification of the polar elements of the EtOAc-soluble fraction of the plant revealed nine novel dammarane-type triterpenes, named cordianols A-I (1-9) along with the known compound cordialin A (10). The structures of these new compounds were elucidated by means of spectral methods including HRFABMS, (1)H NMR, (13)C NMR, and 2D NMR (HMQC, HMBC, NOESY). Absolute configuration at C-23 of compound 7 was determined by an excitone chirality method. Some of these new compounds revealed a hemiketal structure on the A ring and a hydroxylated or epoxidated 20(22)-(E)-ene side chain and showed weak anti-androgenic activity.

Phenolic constituents from Dalbergia cochinchinensis.

Three new phenolic compounds (1-3), along with five known phenolics, 4'-hydroxy-2'-methoxychalcone (4), latinone (5), dalbergiphenol (6), 7-hydroxyflavanone, and dalbergin (7), have been isolated from the stems of Dalbergia cochinchinensis. The structures of 1-3 were established by spectroscopic techniques including 1D and 2D NMR methods. The inhibitory activity against testosterone 5 alpha-reductase, which causes androgen-dependent diseases, was also examined for selected compounds.