Chlorination diversifies Cimicifuga racemosa triterpene glycosides

Extracts from the roots and rhizomes of black cohosh (Cimicifuga racemosa) are widely used as dietary supplements to alleviate menopausal symptoms. State-of-the-art quality control measures involve phytochemical fingerprinting of the triterpene glycosides for species identification and chemical standardization by HPLC. In the course of developing materials and methods for standardization procedures, the major C. racemosa triterpene glycoside (1) was isolated and initially thought to be cimicifugoside (2). Detailed HR-LC-MS and 1D and 2D NMR analysis of 1 and 2 unambiguously revealed that 1 is the chlorine-containing derivative of 2, namely, 25-chlorodeoxycimigenol-3-O-beta-d-xyloside. Accordingly, HPLC profiles of black cohosh preparations require revision of the assignments of the chlorinated (1) and nonchlorinated (2) pair. Besides explaining the substantial shift in polarity (DeltatR[RP-18] ca. 20 min), 25-deoxychlorination opens a new pathway of structural diversification in triterpene glycoside chemistry. As chemical conversion of 2 into 1 could be demonstrated, deoxychlorination may be interpreted as artifact formation. Simultaneously, however, it is a potentially significant pathway for the gastric in vivo conversion ("nature's prodrug") of the relatively polar triterpene glycosides into significantly less polar chlorinated derivatives with altered pharmacological properties.     

Cimiracemosides I-P,new 9,19-cyclolanostane triterpene glycosides from Cimicifuga racemosa

Eight new and 13 known triterpene glycosides, along with the known compounds glyceryl-1-palmitate and daucosterol-6'-linoleate were isolated from the roots/rhizomes of Cimicifuga racemosa. The new compounds, designated as cimiracemosides I-P (1, 3-9), were determined by spectral analysis to be 7-dehydro-23-epi-12,26-dideoxyacteol-3-O-beta-D-xylopyranoside (1), 12-O-acetyl-25-anhydrocimigenol-3-O-alpha-L-arabinopyranoside (3), 12-O-acetyl-25-anhydrocimigenol-3-O-beta-D-xylopyranoside (4), 4',23-O-diacetylshengmanol-3-O-beta-D-xylopyranoside (5), 4',23-O-diacetylshengmanol-3-O-alpha-L-arabinopyranoside (6), 23-epi-acetylacteol-3-O-alpha-L-arabinopyranoside (7), 4'-O-acetyl-26-deoxyactein (8), and 16beta:23;24:25-diepoxy-12beta-O-acetyl-3beta-hydroxy-9,19-cyclolanost-23,26-olide-O- beta-D-xylopyranoside (9).     

Cimipronidine, a cyclic guanidine alkaloid from Cimicifuga racemosa

A new cyclic guanidine alkaloid, cimipronidine (1), together with the known compound fukinolic acid (2), was isolated from the n-BuOH-soluble fraction of Cimicifuga racemosa roots that showed 5-HT7 receptor binding activity. Structure elucidation of 1, a minor constituent, presented unique challenges based on its polarity, but was accomplished with the use of a combination of one- and two-dimensional NMR as well as MS analyses. The relative configuration was established by analyzing the H,H-coupling constants and the results of the 2-D gradient NOESY spectrum. The previously reported serotonergic (5-HT7), highly polar, n-BuOH-soluble fraction was characterized by HPLC-ELSD and was shown to be a mixture containing the following compounds: cimicifugic acids A, B, and F, fukinolic acid, ferulic acid, isoferulic acid, and compound 1, potentially significant as a marker compound of C. racemosa.     

Neocimicigenosides A and B, cycloartane glycosides from the rhizomes of Cimicifuga racemosa and their effects on CRF-stimulated ACTH secretion from AtT-20 cells

Two new cycloartane glycosides, named neocimicigenosides A (1) and B (2), were isolated from the rhizomes of Cimicifuga racemosa. The structures of 1 and 2 were determined on the basis of extensive spectroscopic analysis and enzymatic hydrolysis followed by chromatographic and spectroscopic analyses to be (16S,23R,24S)-24-acetoxy-16,23:16,25-diepoxy-15alpha-hydroxycycloartan-3beta-yl alpha-L-arabinopyranoside (1) and (16S,23R,24S)-24-acetoxy-16,23:16,25-diepoxy-15alpha-hydroxycycloartan-3beta-yl beta-D-xylopyranoside (2), respectively. Neocimicigenosides A and B enhanced CRF-stimulated ACTH secretion from AtT-20 cells.     

Actaeaepoxide 3-O-beta-D-xylopyranoside, a new cycloartane glycoside from the rhizomes of Actaea racemosa (Cimicifuga racemosa)

A new cycloartane glycoside (1) was obtained from a minor triterpene fraction of the rhizome extract of Actaea racemosa (synonym: Cimicifuga racemosa) along with a known compound, cimigenol 3-O-beta-D-xylopyranoside. The structure of 1 was elucidated as 20(S),22(R),23(R),24(S)-12beta-acetoxy-16beta:23,23alpha:24-diepoxy-3beta,22beta,25-trihydroxy-9,19-cyclolanost-7-ene 3-O-beta-D-xylopyranoside (actaeaepoxide 3-O-beta-D-xylopyranoside) on the basis of spectral and chemical evidence.     

Triterpene glycosides from the roots of Astragalus flavescens

Six new triterpene saponins, 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-21-epi-kudzusapogenol A (1), 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-21-epi-kudzusapogenol A (2), 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-22-O-beta-D-glucopyranosyl-21-epi-kudzusapogenol A (3), 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-22-O-beta-D-glucopyranosyl-21-epi-kudzusapogenol A (4), 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-22-O-alpha-L-arabinopyranosyl-21-epi-kudzusapogenol A (5), and 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->2)-beta-D-glucuronopyranosyl-22-O-alpha-L-arabinopyranosyl-21-epi-kudzusapogenol A (6), were isolated from the roots of Astragalus flavescens, together with the known trajanoside B, azukisaponin V, and astragalosides IV, VII, and VIII. Their structures were established mainly by 2D NMR techniques and mass spectrometry.     

Anticomplement activity of cycloartane glycosides from the rhizome of Cimicifuga foetida

A tetranor-cycloartane glycoside and two 9,19-cycloartane glycosides were isolated from the EtOAc-soluble fraction of the rhizome of Cimicifuga foetida. The structures of the compounds were determined to be cimilactone A (1), 25-O-acetylcimigenol 3-O-beta-d-xylopyranoside (2) and cimigenol 3-O-alpha-l-arabinopyranoside (3), respectively, using spectroscopic analysis. The three compounds were examined for their anticomplement activity against the classical pathway of the complement system. Compound 1 showed significant anticomplement activity with an IC(50) value of 28.6 microm, whereas compounds 2 and 3 were inactive.     

Triterpene glycosides from the far eastern sea cucumber Pentamera calcigera II: disulfated glycosides

Three new triterpene glycosides, calcigerosides D(1) (1), D(2) (2), and E (3), have been isolated from the sea cucumber Pentamera calcigera. Their structures have been deduced from extensive spectral analysis (NMR and MS) and chemical evidence. All the compounds are disulfated pentaosides differing in aglycon structure and position of sulfate group, which were determined by the measurement of NT(1) values in the cases of glycosides 1 and 2. Glycoside 1 is a nonholostane derivative, that is, it lacks an 18(20)-lactone, which is very rare among the sea cucumber glycosides.     

Triterpene glycosides from the Far Eastern sea cucumber Cucumaria conicospermium

Four new triterpene glycosides, cucumariosides A(2)-5 (1), A(3)-2 (2), A(3)-3 (3), and isokoreoside A (4), along with the previously isolated koreoside A (5), have been found in the sea cucumber Cucumariaconicospermium. Glycoside 1 was isolated as a native substance, while glycosides 2-5 were identified through their desulfated derivatives. Their structures have been deduced by extensive spectral analysis (NMR and MS) and chemical evidence. All the glycosides contain the same branched pentasaccharide carbohydrate chain but differ in the number and positions of the sulfate groups. Glycoside 1 has one, glycosides 2 and 3 have two, and glycosides 4 and 5 have three sulfate groups. Glycosides 2-5 are non-holostane derivatives; their aglycons lack the 18(20)-lactone and are characterized by shortened side chains, which is a very rare feature among the sea cucumber glycosides.     

Triterpene glycosides from the deep-water North-Pacific sea cucumber Synallactes nozawai Mitsukuri

Five non-sulfated triterpene glycosides, synallactosides A(1) (1), A(2) (2), B(1) (3), B(2) (4), and C (5), have been isolated from the sea cucumber Synallactes nozawai. Their structures have been deduced by extensive analysis of NMR and mass spectra. The glycosides 2-5 are new glycosides. Glycosides 2-4 have carbohydrate chains without precedent in the glycosides from sea cucumbers. This is the first time glycosides are found in members of the family Synallactidae.     

Flavonoid glycosides from Adina racemosa and their inhibitory activities on eukaryotic protein synthesis

From the dried leaves, flowers, and twigs of Adina racemosa, five new flavonoid glycosides, quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-(3-O-trans-p-coumaroyl)-beta-d-galactopyranoside (1), quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O-trans-p-coumaroyl)-alpha-l-rhamnopyranosyl(1-->2)]-(4-O-trans-p-coumaroyl)-beta-d-galactopyranoside (2), kaempferol 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O-trans-p-coumaroyl)-alpha-l-rhamnopyranosyl(1--> 2)]-(4-O-trans-p-coumaroyl)-beta-d-galactopyranoside (3), quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O-trans-p-coumaroyl)-alpha-l-rhamnopyranosyl(1-->2)]-(3-O-trans-p-coumaroyl)-beta-d-galactopyranoside (4), and quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O-trans-caffeoyl)-alpha-l-rhamnopyranosyl(1-->2)]-(3-O-trans-p-coumaroyl)-beta-d-galactopyranoside (5), and eight known compounds were isolated. The structures of the new compounds were determined by spectroscopic and chemical means. Their inhibitory activities on protein synthesis were assessed. The new glycosides were found to be inhibitors of eukayrotic, but not prokaryotic, protein synthesis.     

黑升麻治疗妇女围绝经期综合征的临床研究

目的对黑升麻治疗围绝经期综合征的临床试验进行综述。方法利用国外权威数据库,查找相关的黑升麻临床试验的文献,并对其进行分析。结果研究表明黑升麻能较好地缓解围绝经期的症状。结论黑升麻是治疗围绝经期综合征的具有潜力的药物。     

Triterpene glycosides from the Far-Eastern sea cucumber Pentamera calcigera. 1. Monosulfated glycosides and cytotoxicity of their unsulfated derivatives

Three new monosulfated triterpene glycosides, calcigerosides B (2), C(1) (3), and C(2) (4), along with the known cucumarioside G(2) (1), have been isolated from the sea cucumber Pentamera calcigera. Their structures have been deduced from extensive spectral analysis (NMR and MS) and chemical evidence. Compounds 2-4 present a novel pentasacharide chain never reported before in sea cucumber triterpene glycosides. The desulfated derivatives of calcigerosides B, C(1), and C(2) (5, 7, and 9, respectively) showed moderate cytotoxicity (IC(50) = 5 microg/mL) against a selection of four human and mouse tumor cell lines.     

Triterpene glycosides from the flower petals of sunflower (Helianthus annuus) and their anti-inflammatory activity

Two new oleanane-type triterpene glycosides, named helianthosides 4 (4) and 5 (5), along with four known triterpene glycosides, helianthosides 1 (1), 2 (2), 3 (3), and B (6), were isolated from an n-butanol-soluble fraction of a methanol extract of sunflower (Helianthus annuus) petals. The structures of the two new compounds were determined on the basis of spectroscopic and chemical methods. Upon evaluation of compounds 1-6 for inhibitory activity against 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation (1.7 nmol/ear) in mice, all of the compounds tested exhibited marked anti-inflammatory activity, with ID50 values in the range 65-262 nmol per ear.     

Isolation, structure elucidation, and absolute configuration of 26-deoxyactein from Cimicifuga racemosa and clarification of nomenclature associated with 27-deoxyactein

A new triterpene glycoside, 26-deoxyactein (1), along with two known compounds, 23-epi-26-deoxyactein (2), previously designated as "27-deoxyactein", and actein (3), were isolated from the roots/rhizomes of Cimicifuga racemosa. The structures and absolute stereochemistry of 1 and 2 were established by spectroscopic methods (FABMS, (1)H and (13)C NMR) and single-crystal X-ray data analysis.     

Black cohosh (Cimicifuga racemosa) relaxes the isolated rat thoracic aorta through endothelium-dependent and -independent mechanisms

Aim of the study

The rhizome of the Cimicifuga racemosa (commonly known as black cohosh) has been used in treatment of climacteric complaints for decades in North America and Europe. A number of studies investigated the estrogenic potential of black cohosh, but its effectiveness is still controversial. Recently, it was reported that the extract of black cohosh acted as an agonist at the serotonin (5-HT) receptor and 5-HT derivative was isolated out of the black cohosh extract. Because it is well known that the 5-HT elicited the various cardiovascular effects including vasorelaxation, we investigated the vasorelaxant effects of the extract of black cohosh and its possible mechanisms of action.

Materials and methods

The extract of black cohosh (BcEx) was examined for its vasorelaxant effects in isolated rat aorta. The aortic rings were equilibrated under resting tension and induced reproducible contraction in organ bath. The control contraction was produced by 300 nM NE, and then BcEx were added. In experiments where specific inhibitors were used, they were added 20 min before NE contraction.

Results

BcEx elicited two phases of relaxation in rat aorta pre-contracted with norepinephrine. The first, a rapid relaxation, which occurred within seconds of BcEx administration, was eliminated by pretreatment with N^G-nitro-l-arginine (l-NNA) or methylene blue. The endogenous NO synthase substrate l-Arg markedly reversed the action of l-NNA, indicating that BcEx elicited the vasorelaxant effect via the NO/cGMP pathway. The second, slowly developing relaxation was not affected by the endothelium denudation. BcEx-induced endothelium-independent vasorelaxation appears to involve the inhibition of calcium influx mediated by the opening of inward rectifier potassium channels.

Conclusions

BcEx elicits the vasorelaxant effect via endothelium-dependent and -independent mechanisms and may contribute to a better understanding of a potential link between the use of black cohosh and its beneficial effects on vascular health.     

Absence of positive effect of black cohosh (Cimicifuga racemosa) on fracture healing in osteopenic rodent model

The healing of predominantly metaphyseal fractures in postmenopausal osteoporosis is delayed and comparatively poor. Due to the potential side effects of HRT, natural alternatives are appealing. The aim of this study was to determine whether Cimicifuga racemosa extract BNO 1055 improves metaphyseal fracture healing in severe osteopenic bone in rats. Thirty‐three 12‐week‐old female rats developed severe osteopenia during 10 weeks after ovariectomy. After metaphyseal tibial‐osteotomy and standardized T‐plate‐osteosynthesis, the healing periods in ovariectomized rats (C), 17‐α‐estradiol (E) and Cimicifuga racemosa (CR) supplemented diets were assessed for 35 days. Changes in callus morphology were evaluated qualitatively by biomechanical testing and quantitatively in microradiographies and fluorochrome‐labeled histological sections. The CR‐supplementation slightly improved callus quality and trabecular bone formation. It significantly enhanced the endosteal callus density compared to C group (Cl.Dn.e C: 59.08 ± 21.89, E: 45.95 ± 18.39, CR: 60.85 ± 18.66*), though most of the other morphological parameters examined showed no improvement. The time course of fracture healing did not change due to CR. Estrogen‐supplementation enhanced the biomechanical properties of the fracture site. Trabecular bone was improved indicating the physiological endosteal healing process. The CR‐supplementation did not exhibit positive effects in severe (senile) osteopenic fracture healing as seen in early (postmenopausal) osteoporosis in rats. Callus formation was slightly improved under CR. Estrogen improved fracture healing in severe osteopenic bone, while the extent of callus formation played a minor role. Copyright © 2010 John Wiley & Sons, Ltd.     

Triterpene saponins from clematis mandshurica

Four new triterpene saponins, clematomandshurica saponins A-D (1-4), together with three known saponins (5-7) have been isolated from the roots and rhizomes of Clematis mandshurica. Their structures were elucidated on the basis of their spectroscopic evidence and hydrolysis. Clematomandshurica saponins A and B showed significant inhibitory activity on cyclooxygenase-2 (IC50 = 2.66 and 2.58 microM, respectively).     

Triterpene saponins from Eryngium campestre

Five new triterpene saponins, 3-O-alpha-l-rhamnopyranosyl-(1-->2)-beta-d-glucuronopyranosyl-22-O-beta,beta-dimethylacryloyl-A1-barrigenol (1), 3-O-alpha-l-rhamnopyranosyl-(1-->2)-beta-d-glucuronopyranosyl-22-O-angeloyl-R1-barrigenol (2), 3-O-alpha-l-rhamnopyranosyl-(1-->2)-beta-d-glucuronopyranosyl-21-O-acetyl-22-O-angeloyl-R1-barrigenol (3), 3-O-alpha-l-rhamnopyranosyl-(1-->2)-beta-d-glucuronopyranosyl-21-O-acetyl-22-O-beta,beta-dimethylacryloyl-R1-barrigenol (4), and 3-O-alpha-l-rhamnopyranosyl-(1-->2)-beta-d-glucuronopyranosyl-22-O-angeloyl-28-O-acetyl-R1-barrigenol (5), were isolated from the roots of Eryngium campestre. Their structures were established mainly by 2D NMR techniques and mass spectrometry. Compounds 1-4 and 3-O-beta-d-glucopyranosyl-(1-->2)-[alpha-l-rhamnopyranosyl-(1-->4)]-beta-d-glucuronopyranosyl-22-O-beta,beta-dimethylacryloyl-A1-barrigenol, previously isolated from the same plant, showed a weak cytotoxicity when tested against HCT 116 and HT 29 human colon cancer cells.     

Six secoiridoid glucosides from Adina racemosa

Six novel secoiridoid glucosides, adinosides A (1), B (2), C (3), D (4), E (5), and grandifloroside 11-methyl ester (6) were isolated, together with 27 known compounds, from the dried leaves, flowers, and twigs of Adina racemosa. The structures of the new compounds were determined by spectroscopic (NMR, MS) and chemical means.