Studies on constituents from Chamaecyparis pisifera and antibacterial activity of diterpenes.

In the course of our research for biologically active constituents from coniferous plants, a chromone derivative (1) and an abietane derivative (2) were isolated along with several diterpenes from Chamaecyparis pisifera. Structures of the new compounds were determined to be 5,7-dihydroxy-2-(1-acetyl-2-methoxycarbonylethyl)-chromone and rel-(8R,10R,20S)-8,10,20-trihydroxy-9(10-->20)-abeo-abieta-9,13-dien-12-one by means of spectral methods including two-dimensional NMR experiments. Some of these abietane-type compounds isolated from this plants showed antibacterial activitv against the gram-positive bacteria Staphylococcus aureus and Bacillus subtilis.

Anti-androgenic triterpenoids from the Brazilian medicinal plant, Cordia multispicata.

Compounds 1-6 were isolated from the AcOEt soluble fraction of leaves of the Brazilian medicinal plant, Cordia multispicata, and their structures were elucidated to be 3beta,25-epoxy-21beta-acetoxy-3alpha,22beta-dihydroxyurs-12-en-28-al (1), 3beta,25-epoxy-28-acetoxy-3alpha,21beta,22beta-trihydroxyurs-12-ene (2), 21beta-acetoxy-22beta-hydroxy-3-oxours-12-en-28-al (3), 28-acetoxy-6beta, 21beta,22beta-trihydroxy-3-oxours-12-ene (4), 21beta,22beta-dihydroxy-3-oxours-1 2-en-28-al (5) and 3beta,21beta,22beta-trihydroxyurs-I2-en-28-al (6), respectively, by means of spectral data, especially two dimensional NMR techniques. Triterpenes having the hemiketal structure at the A-ring, an acyloxy group at C-22 and/or ketone at C-3 showed potent anti-androgenic activity.

Gene for a protein capable of enhancing lateral root formation.

Analysis of genes preferentially expressed in hairy roots caused by infection with Agrobacterium rhizogenes has provided insights into the regulation of lateral root formation. A hairy root preferential cDNA, HR7, has been cloned from hairy roots of Hyoscyamus niger. HR7 encodes a novel protein partially homologous to a metallocarboxypeptidase inhibitor and is expressed exclusively in the primordium and base of lateral roots in hairy roots. Overexpression of HR7 in transgenic roots of H. niger dramatically enhances the frequency of lateral root formation. The results of this study indicate that expression of HR7 plays a critical role in initiating lateral root formation.

Antibacterial and antiandrogen flavonoids from Sophora flavescens.

Sixteen flavanones, three flavanonols, and four pterocarpans were isolated from the MeOH extract of the roots of Sophora flavescens. Twelve of these were new compounds, including eight prenylflavanones (1-8), one prenylflavanonol (9) and three novel pterocarpane derivatives (10-12). Their structures were elucidated using NMR and mass spectral methods. Some of these compounds have irregular C10 prenyl moieties at C-8 of the flavanone skeleton. These compounds exhibited significant antibacterial activities against the Gram-positive bacteria Staphylococcus aureus, Bacillus subtilis, S. epidermidis, and Propionibacterium acnes. They also exhibited antiandrogen activities.

Oligosaccharide polyesters from roots of Polygala glomerata.

Seven new sucrose and oligosaccharide esters, glomeratoses A-G, together with four known compounds, 3-O-[(E)-3,4,5-trimethoxycinnamoyl]-beta-D-fructofuranosyl-(2-->1) - (6-O-benzoyl)-alpha-D-glucopyranoside, 3-O-(E)-sinapoyl-beta-D-fructofuranosyl-(2-->1)-[6-O-(E)-sinapo yl]-alpha-D- glucopyranoside, tenuifoliside C and reiniose G were isolated from the roots of Polygala glomerata. Glomeratoses A-G were elucidated as 3-O-[(E)-3,4,5-trimethoxycinnamoyl]-beta-D-fructofuranosyl-(2-->1) -alpha-D- glucopyranoside, 3-O-(E)-sinapoyl-beta-D-fructofuranosyl-(2-->1)-[6-O-(E)-p- coumaroyl]-alpha-D-glucopyranoside, 3-O-[(E)-3,4,5-trimethoxycinnamoyl]-beta-D-fructofuranosyl-(2-->1) -[6-O-(E)- p-coumaroyl]-alpha-D-glucopyranoside, 3-O-[(E)-3,4,5-trimethoxycinnamoyl]-beta-D-fructofuranosyl-(2-->1) -[6-O-(E)- 3,4,5-trimethoxycinnamoyl]-alpha-D-glucopyranoside, 1-O-{6-O-[3-O-(E,E)-(beta, beta'-bis-sinapoyl)-beta-D-fructofuranosyl]}-alpha- D-glucopyranoside intramolecular ester, 1-O-(E)-p-coumaroyl-(3-O-benzoyl)-beta-D- fructofuranosyl-(2-->1)-[beta-D-glucopyranosyl-(1-->2)]-[6-O-acetyl-beta -D- glucopyranoysl-(1-->3)]-[4-O-(E)-feruloyl]-(6-D-acetyl)-alph a-D- glucopyranoside, 1-O-(E)-p-coumaroyl-(3-O-benzoyl)-beta-D-fructofuranosyl-(2-->1)-[ beta-D- glucopyranosyl-(1-->2)]-[6-O-acetyl-beta-D-glucopyranoside-(1-->3)]-{4-O - [4-O-beta-D-glucopyranosyl-(E)-feruloyl]}-[6-O-(E)-p-coumaroyl+ ++]-alpha- D-glucopyranosyl, respectively, on the basis of chemical and spectral evidence.

Polygalasaponins XLII-XLVI from roots of Polygala glomerata.

Five new oleanane-type saponins, polygalasaponins XLII-XLVI, along with two known saponins were isolated from the roots of Polygala glomerata Lour. The structures of polygalasaponins XLII-XLVI were elucidated as 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl- (1-->2)-{4-O-[(E)-3,4-dimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)- alpha-L-rhamnopyranosyl-(1-->2)-[alpha-L-arabinopyranosyl- (1-->3)]-[4-O-(E)-p-methoxycinnamoyl]-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)- alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl- (1-->3)]-{4-O-[(E)-3,4-dimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)- alpha-L-rhamnopyranosyl-(1-->2)-[6-O- acetyl-beta-D-glucopyranosyl-(1-->3)]-{4-O- [(E)-3,4-dimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, 3-O-beta-D- glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)- beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl- (1-->2)-[6-O-acetyl-beta-D-glucopyranosyl-(1-->3)]-{4-O-[(Z)-3, 4-dimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, respectively, on the basis of spectroscopic and chemical evidence.

Nine new triterpene saponins, polygalasaponins XXXIII--XLI from the roots of Polygala fallax Hemsl.

Nine new oleanane-type saponins, polygalasaponins XXXIII--XLI, along with seven known saponins were isolated from the roots of Polygala fallax HEMSL. Polygalasaponins XXXIII-XLI were elucidated as 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl- (1-->2)-(4-O-acetyl)-beta-D-fuco-pyranosyl ester, 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl- (1-->4)-alpha-L-rhamnopyranosyl-(1-->2)-(4-O-acetyl)-beta-D- fucopyranosyl ester, 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)- alpha-L-rhamnopyranosyl-(1-->2)-(3,4-di-O-acetyl)-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)- [(5-O-acetyl)-beta-D-apiofuranosyl-(1-->3)]-alpha-L-rhamnopyranosy l- (1-->2)-(3,4-di-O-acetyl)-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl presenegenin 28-O-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->2)- (3-O-acetyl)-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosyl- (1-->2)-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl- (1-->4)-alpha-L-rhamnopyranosyl-(1-->2)-(4-O-acetyl)-beta-D- fucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl presenegenin 28-O-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->2)- [alpha-L-rhamnopyranosyl-(1-->3)]-(4-O-acetyl)-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)- [beta-D-apiofuranosyl-(1-->3)]-alpha-L-rhamnopyranosyl-(1-->2)- (3,4-di-O-acetyl)-beta-D-fucopyranosyl ester and 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl presenegenin 28-O-beta-D-galactopyranosyl-(1-->4)-beta-D-xylopyranosyl-(1-->4)- [(5-O-acetyl)-beta-D-apiofuranosyl-(-->3)]-alpha-L-rhamnopyranosyl - (1-->2)-(3,4-di-O-acetyl)-beta-D-fucopyranosyl ester, respectively, on the basis of spectroscopic and chemical evidence.

Hypoglycemic effect of extracts from Lagerstroemia speciosa L. leaves in genetically diabetic KK-AY mice.

The hypoglycemic effects of Lagerstroemia speciosa L., known by the Tagalog name of banaba in the Phillipines, were studied using hereditary diabetic mice (Type II, KK-AY/Ta Jcl). The mice were fed a test diet containing 5% of the hot-water extract (HWE) from banaba leaves, 3% of the water eluent of the partial fraction unadsorbed onto HP-20 resin of HWE (HPWE), and 2% of the methanol eluent of the partial fraction adsorbed onto HP-20 resin of it (HPME) for a feeding period of 5 weeks. The elevation of blood plasma glucose level in non-insulin dependent diabetic mice fed the cellulose as control (CEL) diet were almost entirely suppressed by addition of either HWE or HPME in place of cellulose in the CEL diet. Water intakes were inclined to increase gradually in the group fed either CEL or HPWE, but lower in the mice fed either HWE or HPME than in the animals given either CEL or HPME. The level of serum insulin and the amount of urinary excreted glucose were also lowered in mice fed HWE. Plasma total cholesterol level was also lowered in mice fed the either HWE or HPME. It is suggested that HWE, especially HPME, obtained from banaba leaves have beneficial effects on control of the level of plasma glucose in non-insulin dependent diabetes mellitus.

Studies on differentiation inducers. V. Steroid glycosides from periplocae radicis cortex.

Six pregnane glycosides and three cardenolides were isolated as differentiation inducers using mouse myeloid leukemia (Ml) cells from Periplocae Radicis Cortex (bark of Periploca sepium BGE., Asclepiadaceae). The cardenolides showed much higher activities than the pregnane glycosides. Besides these nine compounds, commercially available cardenolides were tested for their differentiation inducing activities using Ml cells. Digitoxin and digoxin induced Ml cells into phagocytic cells, but others did not. In the presence of 1 nM of actinomycin-D, the activity of steroid glycosides was enhanced against Ml cells.

Studies on the constituents of Polygala japonica HOUTT. II. Structures of polygalasaponins XI-XIX.

Nine new oleanane-type saponins polygalasaponins XI-XIX were isolated from the aerial part of Polygala japonica. The structures of these compounds were established on basis of spectroscopic and chemical evidence.

Studies on the constituents of Polygala japonica Houtt. I. Structures of polygalasaponins I-X.

Ten new oleanane-type saponins, polygalasaponins I-X, along with two known saponins, bayogenin-3-O-beta-D-glucopyranoside and lobatoside B were isolated from the aerial part of Polygala japonica Houtt. The structures of these compounds were established on the basis of spectroscopic and chemical evidence.

Cell differentiation-inducing diterpenes from Andrographis paniculata Nees.

The methanol extract of the aerial part of Andrographis paniculata Nees showed potent cell differentiation-inducing activity on mouse myeloid leukemia (M1) cells. From the ethyl acetate-soluble fraction of the methanol extract, six new diterpenoids of ent-labdane type, 14-epi-andrographolide (3), isoandrographolide (4), 14-deoxy-12-methoxyandrographolide (7), 12-epi-14-deoxy-12-methoxyandrographolide (8), 14-deoxy-12-hydroxyandrographolide (9) and 14-deoxy-11-hydroxyandrographolide (10) as well as two new diterpene glucosides, 14-deoxy-11,12-didehydroandrographi-side (12) and 6'-acetylneoandrographolide (14), and four new diterpene dimers, bis-andrograpolides A (15), B (16), C (17) and D (18), were isolated along with six known compounds. The structures of the diterpenoids were determined by means of spectral methods. Some of these compounds showed potent cell differentiation-inducing activity towards M1 cells.

Studies on differentiation inducers. IV. Pregnane derivatives from condurango cortex.

In connection with the study of differentiation inducers from plants, the methanol extract of Condurango Cortex (bark of Marsdenia condurango REICH, Asclepiadaceae) was investigated to examine its differentiation-inducing activity towards mouse myeloid leukemia (M1) cell line. Six pregnane glycosides, including three new compounds, were isolated as differentiation inducers. Each of the six active glycosides has three or four deoxylated sugars which are well-known to occur in Asclepiadaceae plants. M1 cells were differentiated into phagocytic cells by these glycosides, and they were found to be more effective than their aglycones. Condurangoglycosides A (7) and C (8), having a cinnamoyl group in their aglycones, were the most potent differentiation inducers and M1 cells became phagocytic cells after 24 h treatment with these compounds.

Development of a novel method for determination of acetyl-CoA:1-alkyl-sn-glycero-3-phosphocholine acetyltransferase activity and its application to screening for acetyltransferase inhibitors. Inhibition by magnolol and honokiol from Magnoliae cortex.

A method was developed for determining the activity of acetyl-CoA:1-alkyl-sn-glycero-3-phosphocholine acetyltransferase (EC 2.3.1.67), a key enzyme in the biosynthesis of platelet-activating factor (PAF, 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine). The assay involves measurement of the radioactivity in the trichloroacetic acid (TCA)-precipitated complex of radioactive product and albumin after incubation of 1-alkyl-sn-glycero-3-phosphocholine and [3H]acetyl-CoA with rat spleen microsomes or membrane fractions of human polymorphonuclear leukocytes (PMNs). The radioactive product associated with the precipitate was identified as PAF using an ultrahigh-sensitivity TV camera system after extraction and separation by TLC. This TCA method was then used to screen the components of crude preparations that inhibited acetyltransferase activity. Major components from the cortex of Magnoliae (magnolol and honokiol), which have anti-inflammatory and anti-bacterial actions, inhibited the acetyltransferase activity in rat spleen microsomes (IC50, 150 and 150 microM, respectively) and membrane fractions of human PMNs (IC50, 70 and 60 microM, respectively). The inhibitory action of magnolol and honokiol was reversible, and similar to or higher than that of nordihydroguaiaretic acid. PAF production in human PMNs stimulated by the ionophore A23187 was also suppressed dose dependently by magnolol and honokiol. These activities may be relevant to the claimed therapeutic effects of the extract from Magnoliae cortex.

Saponins from Trifolium repens.

From the whole plant of white clover, Trifolium repens, five new triterpenoid saponins, designated cloversaponins I-V, were isolated together with four known saponins, beta-D-glucoronopyranosylsoyasapogenol B, soyasaponin I, soyasaponin II and azukisaponin II as their methyl esters. Their structures were determined by 2H NMR and 13C NMR spectroscopy and chemical evidence.

Triterpenoid saponins from Sophora subprostrata.

From Sophora subprostrata Radix, the roots of Sophora subprostrata, six triterpenoidal saponins having soyasapogenol A, B, sophoradiol and kuzusapogenol A as aglycones, were isolated as their methyl esters. The structure of a new saponin was established to be 3-O-[alpha-L-rhamnopyranosyl(1----2)-D-galactopyranosyl(1----2)-beta-D- glucuronopyranosyl] kuzusapogenol A methyl ester by means of 1H and 13C NMR spectroscopy and chemical evidence.

Studies on differentiation-inducing activities of triterpenes.

Differentiation-inducing activity of over 180 extracts of crude drugs and plants was tested using mouse myeloid leukemia cell line (M1). The methanol extracts of clove (Syzygium aromaticum Merrill et Perry, Myrtaceae) showed remarkable induction of differentiation of M1 cells into macrophage-like cells. From the extract, oleanolic acid (1) and crategolic acid (2) were isolated as the active components. We also tested other triterpenes, such as oleananes, ursanes and dammaranes, to investigate the structure-activity relationship. Some triterpene aglycones showed differentiation-inducing activity, but triterpene glycosides showed little activity. Furthermore, the differentiation-inducing activity of these triterpene compounds was tested against human acute promyelocytic leukemia cell line (HL-60).

[Studies on the anticomplementary activity of Jozann. Anticomplementary activity of steroid derivatives].

Methanol extracts of 53 crude drugs were tested on the anticomplementary activity. Of these, 12 samples showed the potent activity. Of these 12 samples, the extract of Jozann (Dichrome Radix) was studied on the isolation and structural elucidation of the anticomplementary constituents. As active constituents, 3 beta-hydroxystigmast-5-en-7-one was isolated and its structure was confirmed by deriving this compound from beta-sitosterol. The activity of such steroid derivatives as stigmastane, cholestane, androstane and pregnane type steroids were further studied. Of these, several kinds of steroids showed the potent activity. From these experiments, the structure and activity relationship was discussed.

Monoamine oxidase inhibitors from Cinchonae Cortex.

Three strong alkaloidal monoamine oxidase (MAO) inhibitors, quinine (1), cinchonicinol ([ 1S,3'R,4'R]-3-(3-ethenyl-4-piperidinyl)-1-(4-quinolinyl)-1-propanol) (2) and cinchonaminone ([ 3'R,4'S]-2-[2-(3-ethenyl-4-piperidinyl)-acetyl]-1H-indole-3-ethanol) (3), were isolated from Cinchonae Cortex (Cinchona succirubra Pav., Rubiaceae). The structures of 2 and 3 were elucidated on the bases of spectral data and chemical evidence, and 3 is a new alkaloid. The inhibitory effects on MAO of 1, 2, 3 and related alkaloids were assayed. The type of inhibition by 1 with respect to benzylamine as a substrate was competitive.