Hiroko Murata

Molecular Phylogeny of Aristolochia sensu lato (Aristolochiaceae) based on Sequences of rbcL, matK, and phyA Genes, with Special Reference to Differentiation of Chromosome Numbers

Abstract The genus Aristolochia sensu lato contains over 400 species from warm temperate to tropical regions worldwide. Taxonomic treatments of Aristolochia have been ambiguous and controversial. In a recent cladistic analysis based on morphological characters, it was proposed that the genus should be divided into four genera in two subtribes. To reconsider the systematics of Aristolochia sensu lato, we reconstructed its phylogeny based on nucleotide sequences of the chloroplast rbcL gene and the nuclear-encoded phytochrome A (phyA) gene for 19 representative species and the chloroplast matK gene of over 80 species. All phylogenetic trees produced with the three genes indicate that Aristolochia sensu lato is a monophyletic group, consisting of two lineages that correspond to the subtribes Aristolochiinae and Isotrematinae. The matK phylogeny shows that each of the lineages includes two sublineages. The Aristolochiinae clade is composed of the Aristolochia sensu stricto and Pararistolochia clades, and the Isotrematinae clade of the Isotrema and Endodeca clades. Chromosome numbers, including newly reported counts for 30 species, are predominantly congruent with the phylogeny: the Aristolochiinae clade shows chromosome numbers of 2n = 6, 12, 14, or 16, while the Isotrematinae clade is characterized by 2n = 32. In the Isotrematinae clade, the paralogous relationships of the phyA gene suggest that polyploidization might have occurred.

Isolation of apoptosis- and differentiation-inducing substances toward human promyelocytic leukemia HL-60 cells from leaves of Juniperus taxifolia.

A chloroform extract of the leaves of Juniperas taxifolia exhibited a marked antiproliferative effect on human promyelocytic leukemia HL-60 cells at a concentration of 2.5 μg/ml. Deoxypodophyllotoxin (4) was identified in the extract as an outstanding antiproliferative compound, and five diterpenes (1–3, 5, and 6) were isolated as known compounds with weak or no cytotoxicity. These compounds were examined for their respective apoptosis- and differentiation-inducing activities toward HL-60 cells by DNA fragmentation and NBT-reducing assays, respectively. Among them, 7α-hydroxysandaracopimaric acid (6) was found to have a potent differentiation-inducing activity in a dose-dependent manner at 0.125–2 μg/ml (0.39–6.29 μM), together with apoptosis-inducing activity at concentrations of more than 2.5 μg/ml (7.86 μM). Deoxypodophyllotoxin (4) that exerted cytotoxic and apoptosis-inducing activities at 2 ng/ml (5 nM) did not induce differentiation at the same concentration, and the other diterpenes (1–3 and 5) showed no effect on cell differentiation, even at 5 μg/ml. It was thus demonstrated for the first time that 7α-hydroxysandaracopimaric acid was an effective differentiation-inducing compound toward HL-60 cells.

Stilbene derivatives in the stem of parthenocissus quinquefolia

From the stem of Parthenocissus quinquefolia, two new resveratrol oligomers, parthenocissins A and B, were isolated in addition to three known stilbenes. The structures, 1Z-2α,3β-2-(3,5-dihydroxyphenyl)-2,3-dihydro-3-(4-hydroxyphenyl)-1-[(4-hydroxyphenyl)methylene]-1H-indene-4,6-diol (parthenocissin A) and 1Z,2α,3β,4′α,5′β - 2 - (3,5 - dihydroxyphenyl) - 2,3 - dihydro - 3(4 - hydroxyphenyl) - 1 - [{4′ - (3,5 - dihydroxyphenyl) - 3′ - (4-hydroxyphenyl)-4,5-dihydrofurano(2′,3′ : 4,3)-phenyl}methylene]-1H-indene-4,6-diol (parthenocissin B), were elucidated by the analysis of spectral data including 2D-NMR and NOE experiment.

Flavonol glycosides in the roots of Epimedium diphyllum

Two new 8-y,y-dimethylallylkaempferol glycosides, diphyllosldes A and B, were isolated from the roots and rhizomes of Eptmedium diphyllum m addition to four known flavonol glycosldes Their structures were established by spectroscopic methods. The difference m chemical constituents between aerial and underground parts of the plant is also described INTRODUCTION In Eplmedium species, the aerial parts are mainly utilized as tonic, robust agents, whereas the underground parts are used for treating asthamatlc fits and menstrual irregularity. In recent studies of the aerial parts of the plant, 12 flavonol glycosldes have been found [l-6]. The constituents of the underground parts (roots and rhizomes) of E. diphyllum (Morr. et Decne) Lodd. have now been investigated. RESULTS AND DISCUSSION The butanol-soluble portion of the 70% methanolic extract was separated by use of medium pressure liquid chromatography on silica gel to give seven compounds (l-7). Compound 1, mp 197-199”, obtained as a pale yellow powder, was a flavonol glycoside. The aglycone moiety was deduced to be 8-y,y-dlmethylallylkaempferol (noranhydroicaritm) by the fragment m/z 354 (CZ0Hi80s) in the EI mass spectrum, which was supported by the ‘H NMR spectral data as follows; a one-proton singlet (6.64 ppm) assignable to the proton at C-6 because of a glycosylation at C-7 [S, 61 and two two-protons doublets (6.96 and 7 86 ppm, J=8.8 Hz) those of C-3’,5’ and C-2’@ of kaempferol, the presence of y,y-dimethylallyl group substituted at C-8 was confirmed by the peaks of two threeprotons singlets at 1.63 and 1.71 ppm (-CMe,), a twoprotons multiplet at 3 04ppm (C&CH=C), a oneproton triplet at 5.2 ppm (J = 6 2 Hz) (CH,-Cg=C <). In the ‘H NMR spectrum, the conspicuous dfirences of 1 compared with sagittatoside A (anhydroicaritm 3-0p-D-ghCOSyl-( l-2)-a+rhamnoside [6] were that the chemical shift of H-6 shifted towards downfield by 0.3 ppm and no peak due to methoxyl group was observed. The data showed that sugars are substituted at both C-3 and at C-7 of noranhydroicantin. The UV spectrum also supported the location of the sugars, since bathochromic shifts indicated that the C-S and C-4 hydroxyls were unsubstltuted.

Cycloartane triterpenoids from Aglaia harmsiana

Abstract Two new and two known cycloartane-type triterpenoids were isolated from the leaves of Aglaia harmsiana. The structures were determined using 1H, 13C and 2D NMR techniques.

Pregnanes and triterpenoid hydroperoxides from the leaves of Aglaia grandis

Abstract Three pregnanes and two known cycloartane-type triterpenoid hydroperoxides were isolated from the leaves of Aglaia grandis . Their structures were determined using 1 H, 13 C and 2D NMR techniques.

New Neolignan and Phenylpropanoid Glycosides in Juniperus communis var. depressa

Two new neolignan glucosides (junipercomnosides C and D) and two new phenypropanoid glycosides (junipercomnosides E and F) were isolated from aerial parts of Juniperus communis var. depressa along with seven known phenylpropanoid glycosides. The structures of the isolated compounds were determined by spectral analysis, in particular by the detailed analysis of 2D NMR and CD spectra.

Eryvarins F and G, two 3-phenoxychromones from the roots of Erythrina variegata

Two 3-phenoxychromones, eryvarins F and G, were isolated from the roots of Erythrina variegata. Their structures were established to be 3-(2,4-dihydroxyphenoxy)-7-hydroxy-6,8-di(3,3-dimethylallyl)chromen-4-one and 3-(2,4-dihydroxyphenoxy)-8-(3,3-dimethylallyl)-2,2-dimethylpyrano[5,6:6,7]chromen-4-one on the basis of spectroscopic and chemical evidence. Eryvarins F and G are unusual 3-phenoxychromone derivatives with two isoprenoid groups.

Two flavonol glycosides from the underground parts of Vancouveria hexandra.

Two new flavonol glycosides were isolated from the roots and rhizomes of Vancouveria hexandra. The glycosides were determined by chemical and spectral means to be anhydroicaritin 3-galactosyl(1----3) rhamnoside-7-glucoside and anhydroicaritin 3-[6-O-acetyl-galactosyl(1----3)rhamnoside]-7-glucoside.

Phenolic constituents from seeds of Coptis japonica var. dissecta

Abstract In addition to coumarinolignans (cleomiscosin A aquillochin) and 7,4′-dihydroxy-5-methoxyflavanone, a new dihydrochalcone, 2′,4,4′-trihydroxy-6′-methoxydihydrochalcone, was isolated from the seeds of Coptis japonica var. dissecta. The structure of the dihydrochalcone was confirmed by comparison with relevant synthetic samples.