Studies on the constituents of Broussonetia species X. Six new alkaloids from Broussonetia kazinoki Sieb.

Six new alkaloids, broussonetines W, X, M1, U1, J3, and J2 (1-6) were isolated from the branches of Broussonetia kazinoki SIEB. (Moraceae) as minor constituents. They were formulated as (2R,3R,4R,5R)-2-hydroxy-methyl-3,4-dihydroxy-5-17-(cyclohexy-2-on-1(6)-enyl)heptyllpyrrolidine (1), (2R,3S,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-17-(cyclohexy-2-on-1(6)-enyl)heptyl]pyrrolidine-4-O-beta-D-glucopyranoside (2), (2R,3R,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(9R)-9,13-dihydroxytridecyl]- pyrrolidine (3), (2S,3S,4S)-2-hydroxymethyl-3,4-dihydroxy-5-(10-oxo-13-hydroxytridecyl)-5- pyrroline (4), (2R)-2-[(IS,2S)-1,2-dihydroxy-8-1(2R,3R,4R,5R)-5-(2-hydroxymethyl-3,4-dihydroxy-1-acetylpyrrolidinyl)loctyl]piperidine (5), (2R)-2-[(1S,2S)-1,2-dihydroxy-8-[(2R,3R, 4R,5R)-5-(2-hydroxymethy]-3,4-dihydroxypyrrolidinyl)]octyl]piperidine (6).

Two new pyrrolidine alkaloids, radicamines A and B, as inhibitors of alpha-glucosidase from Lobelia chinensis Lour.

Two new pyrrolidine alkaloids, radicamines A and B were isolated as inhibitors of alpha-glucosidase from Lobelia chinensis Lour. (Campanulaceae). Radicamines A and B were formulated as (2S,3S,4S,5S)-2-hydroxymethyl-3,4-dihydroxy-5-(3-hydroxy-4-methoxyphenyl)-pyrrolidine (1) and (2S,3S,4S,5S)-2-hydroxymethyl-3,4-dihydroxy-5-(4-hydroxyphenyl)-pyrrolidine (2) on the basis of spectroscopic analyses and chemical methods.

Effects of fukinolic acid and cimicifugic acids from Cimicifuga species on collagenolytic activity.

The inhibitory collagenolytic activity (47-64% inhibition in 0.22-0.24 microM) of fukinolic acid and cimicifugic acids A, B, and C, which are esters of fukiic acid (3',4'-dihydroxybenzyl tartaric acid) was more potent than that (20-37% inhibition in 0.23-0.24 microM) of cimicifugic acids D, E, F, which are esters of pscidic acid (4'-hydroxybenzyl tartaric acid). Since fukiic acid showed weaker inhibition, and caffeic acid, ferulic acid, isoferulic acid, and p-coumaric acid showed far weaker activities, the entire structures of fukinolic acid and cimicifugic acids A, B, and C proved to be responsible for the inhibitory activities. Trypsin and pronase E hydrolyzed collagen nonselectively alone or in addition to collagenase. These collagenolytic activities were also inhibited by fukinolic acid. These results show that fukinolic acid may inhibit either the collagenolytic activities specific to collagenase or nonspecific to other emzymes. The present studies suggest the potential effect of fukinolic acid and cimicifugic acids of Cimicifuga rhizomes in preventing collagen degradation by collagenases or collagenolytic enzymes under pathological conditions, wound healing, or inflammation.

[Studies on the constituents of Cimicifuga species].

The chemical constituents of Cimicifuga simplex and its related species were reexamined using HPLC and high resolution spectral analysis. From C. simplex, C. acerina and C. japonica, a new alkaloid, 59 new cycloartane triterpene glycosides and 11 new aromatic constituents were isolated with the previously reported compounds. The latter aromatic constituents were identified in HPLC analysis of the extracts from crude drugs due to C. dahurica, C. heracleifolia and C. foetida. Several interesting topics in this study are reviewed about separation of unstable and closely related compounds by HPLC, rearrangement reactions, chemical conversion for confirmation of absolute configuration, isomerism in NMR solutions, X-ray crystal analysis, spectral analysis of complicated structures, isolation of glycoside malonates, high yield of enzymatic hydrolysis, specific CD curve due to a cycloart-7-ene system, and so on. Biological activities of a major glycoside, cimicifugoside, a modified triterpene, acerinol, and benzyltartaric acid derivatives such as fukinolic acid and cimicifugic acids were also mentioned.

Studies on the constituents of Cimicifuga species. XXVIII. Cycloart-7-enol glycosides from the underground parts of Cimicifuga simplex Wormsk.

Four new cycloart-7-ene triterpenol arabinosides, bugbanosides C-F, were isolated from the underground parts of Cimicifuga simplex Wormsk. (Ranunculaceae). The structures were elucidated as 12beta-acetoxy-3beta,15alpha,-24R,25-tetrahydroxy-16,23-dione-cycloart-7-ene 3-O-alpha-arabinopyranoside, 12beta-acetoxy-24R,25-epoxy-3beta,15alpha-dihydroxy-16,23-dione-cycloart-7-ene 3-O-alpha-L-arabinopyranoside, 12beta-acetoxy-24R,25-epoxy-3beta-hydroxy-16,23-dione-cycloart-7-ene 3-O-alpha-L-arabinopyranoside, and 16,23R:16,24S-diepoxy-3beta,12beta,15alpha,25-tetrahydroxy-cycloart-7-ene 3-O-alpha-L-arabinopyranoside on the basis of spectral and chemical evidence. The circular dichroism (CD) of bugbanosides C-F showed strong negative maxima at 214-217 nm due to a cycloart-7-ene system, as well as other cycloart-7-ene triterpenes. The CD data showed to be useful in determining basic skeletons, including absolute stereostructures of cycloart-7-ene triterpenes.

Studies on the constituents of Broussonetia species VIII. Four new pyrrolidine alkaloids, broussonetines R, S, T, and V and a new pyrroline alkaloid, broussonetine U, from Broussonetia kazinoki Sieb.

Four new pyrrolidine alkaloids, broussonetines R, S, T, and V and a new pyrroline alkaloid, broussonetine U were isolated from the branches of Broussonetia kazinoki SIEB. (Moraceae) in low yield. Broussonetines R, S and T were formulated as (2R,3R,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(1R)-1-hydroxy-3-[6-(4-hydroxybutyl)-cyclohexy-2-on-1(6)-enyllpropyl] pyrrolidine (1), (2R,3R,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(1R,10S)-1,10,13-trihydroxytridecyl] pyrrolidine (2), (2R,3R,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(1R,5S)-1,5, 13-trihydroxy-10-oxo-tridecyl] pyrrolidine (3). And broussonetines U and V were proposed to be (2S,3S,4S)-2-hydroxymethyl-3, 4-dihydroxy-5-(9-oxo-13-hydroxytridecyl)-5-pyrroline (4), (2R,3S,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(E)-9-oxo-13-hydroxy-3-tridecenyl] pyrrolidine (5), respectively, by spectroscopic and chemical methods.

The biosynthesis of broussonetines: origin of the carbon skeleton.

Broussonetines are glycosidase-inhibitory alkaloids obtained from Broussonetia kazinoki. Feeding experiments using [1-13C]glucose and 13C-NMR spectroscopic studies showed that broussonetines are biosynthesized through routes similar to those of sphingosine and phytosphingosine.

Studies on the constituents of Broussonetia species. VII. Four new pyrrolidine alkaloids, broussonetines M, O, P, and Q, as inhibitors of glycosidase, from Broussonetia kazinoki SIEB.

Four new pyrrolidine alkaloids, broussonetines M, O, P, and Q, were isolated from the branches of Broussonetia kazinoki SIEB, (Moraceae). Broussonetines M, O, P, and Q were formulated as (2R,3R,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(10S)-10,13-dihydroxy-tri decyl]pyrrolidine (1), (2R,3R,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(E)9-oxo-13-hydroxy-3- tridecenyl]pyrrolidine (2), (2R,3R,4R,5R)-2-hydroxymethyl-3,4-dihydroxy-5-[(E)10-oxo-13-hydroxy-3-++ +tridecenyl]pyrrolidine (3), and (2R,3S,4R,5R)-2-hydroxymethyl-3-hydroxy-4-(beta-D-glucopyranosyloxy++ +)-5-[10-oxo-13-(beta-D-glucopyranosyloxy)tridecyl]pyrrolidine (4) respectively, by spectroscopic and chemical methods. 1-4 inhibited beta-glucosidase, beta-galactosidase and beta-mannosidase.

Phylogenetic relationship of six Glycyrrhiza species based on rbcL sequences and chemical constituents.

The nucleotide sequences of ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit gene (rbcL) of Glycyrrhiza glabra, G. uralensis, G. inflata, G. echinata, G. macedonica and G. pallidiflora have been determined to construct their phylogenetic tree. Based on these sequences, the six Glycyrrhiza species were divided into two groups: three, G. glabra, G. uralensis, and G. inflata, which produce glycyrrhizin as a major saponin, and the others, G. echinata, G. macedonica and G. pallidiflora, which produce macedonoside C as a major saponin. Among the three glycyrrhizin-producing species, only two nucleotide substitutions were observed between the rbcL sequences of G. glabra and G. uralensis, and the sequence of G. uralensis was identical to that of G. inflata, indicating that G. uralensis and G. inflata are closely related. Among the three macedonoside C-producing species, only one nucleotide substitution was observed between those of G. echinata and G. macedonica, indicating that these two species are also closely related.

Vasoactive effects of cimicifugic acids C and D, and fukinolic acid in cimicifuga rhizome.

Vasoactive effects of cimicifugic acids A-E, fukinolic acid and fukiic acid isolated from Cimicifuga plants were investigated using rat aortic strips. Cimicifugic acid D and fukinolic acid at 3x10(-4) M caused a sustained, slowly developing relaxation of aortic strips precontracted with norepinephrine (NE) in preparations with or without endothelium. Cimicifugic acid C inversely caused a weak contraction. Cimicifugic acids A, B and E and fukiic acid showed no vasoactivity at 3x10(-4) M. Cimicifugic acids A-E and fukinolic acid are esters between cinnamic acids and the hydroxyl group of benzyltartaric acids. For the manifestation of vasoactivity in the rat aorta, it is concluded that in the cinnamic acid moiety, a caffeoyl group might be necessary for the relaxation activity, and the p-coumaroyl group causes contraction. Concentration response curves for the Ca2+-induced contracture of depolarized aortic strips with isotonic high K+ were not affected by cimicifugic acid D or fukinolic acid. The Ca2+-induced contraction of aortic strips, preincubated with 10(-6) M NE in the presence of 10(-6) M nicardipine and 0.01 mM EGTA in Ca2+-free solution, were inhibited by cimicifugic acid D and fukinolic acid. These results indicated that the inhibition by cimicifugic acid D and fukinolic acid of the NE-induced contraction of rat aorta are attributable to the suppression of Ca2+ influx from the extracellular space enhanced by NE.

Studies on inhibitory activities of fukiic acid esters on germination, alpha-amylase and carboxypeptidase A.

Fukinolic acid (1) and cimicifugic acid A (2), caffeic acid analogs, as well as rosmarinic acid (3) and caffeic acid (4) showed inhibition on seed germination and seedling growth. The potency of 1 and 2 was comparable with that of 3. Compounds 1 and 2 also showed strong inhibitory activities as well as 3 and 4 on alpha-amylase. The activity of 1 was higher than that of acarbose used as a positive control, and its 50% inhibitory concentration (IC50) was 2.41 x 10(-5) M. Compounds 1 and 2 also showed inhibitory activities strong as 3 and stronger than 4 on carboxypeptidase A. The activities of 1 and 2 were higher than that of 1, 10-phenanthroline used as a positive control.

Antimalarial activity and nucleoside transport inhibitory activity of the triterpenic constituents of Cimicifuga spp.

The in vitro antimalarial activity against human malaria parasite (Plasmodium falciparum, FCR-3 strain) was examined using 59 triterpenoids obtained during studies on the triterpenic constituents of Cimicifuga spp. The 50% effective concentration values (EC50) of 25 active triterpenoids were 1.0-3.0 microM, and 19 of the compounds had a common 16, 23:23, 26:24, 25-triepoxy group in the side-chain moieties. Among the active triterpenoids, 9 also showed significant inhibition of nucleoside transport in mouse splenocytes. A relationship between the antimalarial activity and the inhibition of nucleoside transport involving these triterpenoids is discussed.

Studies on the constituents of Broussonetia species. III. Two new pyrrolidine alkaloids, broussonetines G and H, as inhibitors of glycosidase, from Broussonetia kazinoki Sieb.

Two new pyrrolidine alkaloids, broussonetines G and H, were isolated from the branches of Broussonetia kazinoki SIEB. (Moraceae). Broussonetines G and H were formulated as 2 beta-hydroxymethyl-3 alpha, 4 beta-dihydroxy-5 alpha-(1-hydroxy- 6:10;10:13-diepoxytridecyl)-pyrrolidine (1) and 2 beta-hydroxymethyl-3 alpha, 4 beta-dihydroxy-5 alpha-(1-hydroxy- 5:9;9:13-diepoxytridecyl)-pyrrolidine (2), respectively, by spectroscopic methods. 1 and 2 inhibited beta-glucosidase, beta-galactosidase and beta-mannosidase.

Studies on the constituents of Broussonetia species. II. Six new pyrrolidine alkaloids, broussonetine A, B, E, F and broussonetinine A and B, as inhibitors of glycosidases from Broussonetia kazinoki Sieb.

Six new pyrrolidine alkaloids called broussonetine A, B, E, F, and broussonetinine A and B were isolated from the branches of Broussonetia kazinoki Sieb. (Moraceae). Broussonetine A, B, E and F were formulated as 2 beta-hydroxymethyl-3 beta-hydroxy-5-alpha- (10-oxo-13-hydroxytridecyl)-pyrrolidine-4-O-beta-D-glucopyranoside (1), 2 beta-hydroxymethyl-3 beta-hydroxy-5 alpha-(9-oxo-13-hydroxytridecyl)-pyrrolidine-4-O-beta-D-glucopy ran oside (2), 2 beta-hydroxymethyl-3 alpha,4 beta-dihydroxy-5 alpha-(1,13-dihydroxy-10-oxo-tridecyl)-pyrrolidine (3), and 2 beta-hydroxymethyl-3 alpha,4 beta-dihydroxy-5 alpha-(1,13-dihydroxy-9-oxo-tridecyl)-pyrrolidine (4), respectively. Broussonetinine A and B (5 and 6) were also isolated and identified as the aglycones of 1 and 2. 3 and 4 exhibited a strong inhibition of alpha-glucosidase, beta-glucosidase, beta-galactosidase and beta-mannosidase, while 5 and 6 showed a strong inhibition of beta-galactosidase and alpha-mannosidase.

Saikosaponin homologues from Verbascum spp. The structures of mulleinsaponins I-VII.

From the aerial parts of Verbascum (V.) sinaiticum, V. thapsiforme, V. fruticulosum and Celsia roripifolia, seven new saikosaponin homologues, called mulleinsaponins I-VII, having 13,28-epoxy-olean-11-ene skelton were isolated together with eight known saikosaponin homologues, 3-O-beta-D-fucopyranosyl saikogenin F, saikosaponin a, desrhamnosylverbascosaponin, songarosaponins C, D, mimengoside A and buddlejasaponins I, IV. The structures of mulleinsaponins I-VII were characterized as 3-O-beta-D-glucopyronosyl-(1-->3)-beta-D-fucopyranosyl-6-deoxy- saikogenin F, 3-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->3)-beta-D - fucopyranosyl-16-deoxysaikogenin F, 3-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->3)-beta-D - fucopyranosyl-saikogenin F, 3-O-alpha-L-rhamnopyranosyl- (1-->4)-beta-D-glucopyranosyl-(1-->3)-[beta-D-glucopyranosyl-(1-->2)]-be ta -D-fucopyranosyl-21 beta-hydroxysaikogenin F, 3-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->3)-[beta- D- glucopyranosyl-(1-->2)]-beta-D-fucopyranosyl-21 beta-acetoxysaikogenin F, 3-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D- glucopyranosyl-(1-->3)-[beta-D-glucopyranosyl-(1-->2)]-beta-D- fucopyranosyl-16 beta-acetoxysaikogenin F and 3-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl- (1-->3)-[beta-D-glucopyranosyl-(1-->2)]-beta-D-fucopyranosylsaikogeni n F 16-O-beta-D-glucopyranoside, respectively, from chemical and spectroscopic evidence.

Studies on the constituents of Cimicifuga species. XIX. Eight new glycosides from Cimicifuga simplex Wormsk.

Eight new glycosides were isolated from Cimicifuga simplex (Ranunculaceae), and their structures were determined to be 23-O-acetyl-7-8-didehydroshengmanol-3-O-alpha-L-arabinopyranosi de (1), 24-epi-24-O-acetyl-7,-8- didehydrohydroshengmanol-3-O-beta-D-galactopyranoside (2), 7,8-didehydrocimigenol-3-O-beta-D-galacytopyranoside (3), 24-epi-24-O-acetylhydroshengmanol-3-O-beta-D-galactopyranoside (4), cimigenol-3-O-beta-D-galactopyranoside (5), 25-O-methylcimigenol-3-O-beta-D-galactopyranoside (6), 25-O-acetylcimigenol-3-O-beta-D-galactopyranoside (7) and 25-O-acetylcimigenol-3-O-beta-D-glucopyranoside (8). Genuine aglycones were obtained by the hydrolysis of 1--7 with lactase F[Amano] and of 8 with cellulase T[amano]4. Acerinol was prepared from 7,8-didehydrocimigenol and showed antilipemic effects.

[The medicine chest handed down in the Takaya family, doctors of Sendai-han].

There is a medicine chest with design of Chrysanthemums in Makie made for Edo-era and handed down in the Takaya family, doctors of Sendai-han. Many kinds of crude drugs, named by one Chinese character, are kept in this chest. They were classified into three types, namely, [Japanese characters]. Identification of these crude drugs was carried out and the meaning of the classification was brought up as a question.

Phytogrowth-inhibitory activities of sulfur-containing compounds. II. The inhibitory activities of thiosalicylic acid and dihydro-2(3H)-thiophenone-related compounds on plant growth.

Thiosalicylic acid (I) showed rather strong inhibitory activity on the growth of roots of all plants treated except Abelmoschus esculentus Moench at the concentration of 5.0 x 10(-4) M. This compound strongly inhibited the growth of the root of Echinochloa utilis Ohwi et Yabuno even at the low concentration of 5.0 x 10(-5) M. Dihydro-2(3H)-thiophenone (VII) also exhibited inhibitory activity on the growth of roots of all plants treated except Glycine max Merrill. Both compounds inhibited the germination of seeds of some plants at the concentration of 1.0 x 10(-3) M. In I-related compounds (I-V), methyl acetylthiosalicylate (IV) had the strongest inhibitory activity, while in VII-related compounds (VII-XI), 4-hydroxy-2(5H)-thiophenone (VIII) showed the most potent inhibitory activity. The amount of chlorophyll in the cotyledon of Brassica campestris L. subsp. rapa Hook. f. et Anders treated with all compounds except tetrahydrothiophene (XI) was lower than that of the control group.

5-Lipoxygenase inhibitors isolated from the mushroom Boletopsis leucomelas (Pers.) Fayod.

Terphenyl compounds, tentatively named Bl-I (1), Bl-II (2), Bl-III (3), Bl-IV (4) and Bl-V (5), showing 5-lipoxygenase inhibitory activity have been isolated from the mushroom Boletopsis leucomelas (Pers.) Fayod. On the basis of physico-chemical and spectral evidence, they were concluded to be a series of cycloleucomelone-leucoacetates.

Isolation of cytotoxic substance, kalafungin from an alkalophilic actinomycete, Nocardiopsis dassonvillei subsp. prasina.

An alkalophilic actinomycete, strain OPC-553 regarded as Nocardiopsis dassonvillei subsp. prasina, produced the cytotoxic substance, TS-1, which showed a marked inhibitory activity against L5178Y mouse leukemic cell in vitro. The cytotoxicity of TS-1 on this cell was very strong and its ID50 was 0.018 micrograms/ml. Through direct comparison of its spectral data with those of an authentic sample, TS-1 was identified as the antifungal antibiotic, kalafungin, already isolated from the culture broth of Streptomyces tanashiensis. However, the isolation of kalafungin from an alkalophilic actinomycete and its cytotoxicity are reported for the first time in this paper.