Three New C_(21) Steroidal Glycosides from the Roots of Cynanchum komarovii Al.Iljinski

Three new C21 steroidal glycosides named komaroside A, komaroside B, komaroside C were isolated from the ethanolic extract of the roots of Cynanchum komarovii Al.Iljinski (Asclepiadaceae), their structures were determined by physiochemical and spectroscopic analysis.

Two New C-21 Steroidal Glycosides from Cynanchum aurichulatum

Two new C21 steroidal glycosides, cynanauriculoside I and cynanauriculoside II, were isolated from the roots of Cynanchum aurichulatum. Their structures were established using spectroscopic methods including one and two-dimensional NMR.

New C(21) steroidal glycoside from Cynanchum paniculatum

A new C21 steroidal glycoside, neocynapanogenin F 3-O-β-D-thevetoside 1, as well as its known aglycone neocynapanogenin F 2, which have an aberrant 13, 14：14, 15-disecopregnane-type skeleton were isolated from the root of Cynanchum paniculatum. Their structures were elucidated on the basis of spectroscopic data analysis. These compounds showed certain cytotoxic activities in vitro.

Three new C_(21) steroidal glycosides from the roots of Cynanchum inamoenum

Three new C21 steroidal glycosides named inamoside E （1）, inamoside F （2） and inamoside G （3） were isolated from the roots of Cynanchum inamoenum （Maxim.） Loes. Their structures were determined by spectroscopic analysis, especially by ID and 2D NMR experiments.

A new C_(21)-steroidal glycoside from Cynanchum stauntonii

A new C21-steroidal glycoside with two known compounds were isolated from the root of Cynanchum Stauntonii. Based on the spectral analysis, including MS, 1H NMR, 13C NMR, DEPT, 1H-1H COSY, 13C-1H COSY, HMQC and HMBC, their chemical structures were determinated as glaucogenin-C 3-O-α-L-cymaropyranosyl-（1→ 4）-β-o-digitoxopyranosyl-（1→4）-β-D-canaropyranoside （1）, stigmasterol （2） and ursolic acid （3）.

A Steroidal Glycoside from Cynanchum versicolor Bunge

A new C21 steroidal glycoside, named cynanversicoside F （1）, was isolated from the root of Cynanchum versicolor Bunge. Its structure was established as glaucogenin-A 3-O-β-D- digitoxopyranosyl-（1→4）-β-D-cymaropyranoside by spectroscopic and chemical methods.

Two New C21 Steroidal Glycosides from the Roots of Cynanchum paniculatum

Two new C21 steroidal glycosides,paniculatumosides H and I,together with four known ones were isolated from the roots of Cynanchum paniculatum(Bge.)Kitag.Their structures were identifed by spectroscopic methods including extensive 1D and 2D NMR techniques.All compounds were subjected to detect the anti-tobacco mosaic virus(TMV)activities and their cytotoxities against three human tumor cell lines(SMMC-7721,MDA-MB-231 and A549).The results showed that compounds 1 and 5 exhibited potent protective activities against TMV,while 2,4 and 6 had moderate efects on the SMMC7721 cancer cells viability.

白首乌(Cynanchum bungi Decne)脱分化诱导与内源IAA和核酸变化

外源生长素对白首乌脱分化是必要的，效果最好的是２ ，４ － Ｄ，浓度范围０ ．１ ～０ ．３ｍｇ／ Ｌ；白首乌脱分化的起动和细胞旺盛分裂伴随着内源 Ｉ Ａ Ａ 含量的迅速提高， Ｔｒｐ 含量的变化与 Ｉ Ａ Ａ 相似，但其峰值比 Ｉ Ａ Ａ 提前，这为旺盛分裂期 Ｉ Ａ Ａ 的大量形成奠定基础； Ｄ Ｎ Ａ 和 Ｒ Ｎ Ａ 含量的峰值也出现在旺盛分裂期，这与此时期 Ｉ Ａ Ａ 的大量及时形成和旺盛的代谢活动有关，从形成期直至衰老，内源 Ｉ Ａ Ａ、 Ｔｒｐ 和 Ｄ Ｎ Ａ、 Ｒ Ｎ Ａ 含量下降并保持较低水平。

THE MECHANISM OF ANTITUMOR ACTIVITY OF TOTAL GLUCOSIDES EXTRACTED FROM CYNANCHUM AURICULA-TUM ROYLE (CA)

Since the chemical structure of total glucosides from Cynanchum Auriculatum Royle (CA) is similar to that of the steroline of Marsdenia Condurago Reich, a compound which exhibits antitumor activity, research into the antitumor activity of CA was carried out. Its mechanism of action was studied in vivo with C 57BL/6 mice bearing Lewis lung carcinoma and in vitro, with two mouse tumor cell lines: S180 and EAC. CA inhibited to a certain extent the growth of subcutaneously inoculated Lewis lung carcinoma and its pulmonary metastasis, and augmented the antitumor effect of cyclophosphamide. It showed a killing effect on the EAC and S180 tumor cells of mice in vitro as well. It blocked the tumor cells of solid Lewis lung carcinoma from entering into the S stage from G1 and inhibited DNA synthesis of S180 and EAC tumor cells of mice in vitro. It also markedly increased the number of mononuclear Mφ of tumor bearing mice, stimulated the macrophagic activity of their intraperitoneal Mφ, raised the percentage of ANAE(+) lymphocytes in peripheral blood and enhanced the ABC reaction and antibody formation in tumor bearing mice.

Separation and Purification of Acetophenones from Cynanchum bengei Decne Root Bark by Combination of Silica Gel and High-speed Counter-current Chromatography

[Objectives] To develop a method for separation and purification of acetophenones from Cynanchum bengei Decne root bark by combination of silica gel and high-speed counter-current chromatography( HSCCC). [Methods]The crude extract of Cynanchum bengei Decne root bark was separated by silica gel column chromatography,and parts A and B containing acetophenones were obtained. Then,parts A and B were separated by HSCCC with a two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-water( 4∶ 6∶ 4. 5∶ 5. 5 and4∶ 6 ∶ 3 ∶ 7, V/V), respectively. [Results] From 260 mg of part A, four compounds with p-dihydroxybenzene 3. 9 mg(Ⅰ),4-hydroxyacetophenone 17. 1 mg( Ⅱ),2,5-di-hydroxyacetophenone 13. 3 mg(Ⅲ) and 2,4-dihydroxyaceto-phenone 21. 0 mg(Ⅳ) were obtained. And from 300 mg of part B,136 mg of Radix Cynanchi Bungei benzophenone(Ⅴ) was obtained. The purity of compounds determined by HPLC was 97. 0%,96. 6%,99. 2%,99. 7%,99. 5%,respectively. [Conclusions] The established method is simple and efficient. It can be used for separation of acetophenones from Cynanchum bengei Decne root bark and has better practical value,which could provide a reference basis for development and utilization of Cynanchum bengei Decne root bark.

Protective effects of Radix Cynanchum bungei on ischemia-induced neuronal and cognitive impairment in mice

OBJECTIVE To investigate the effects of Radix Cynanchum bungei extract(RCBE) on cerebral ischemia-reperfusion(I/R) and acute cerebral ischemia induced impairment in mice.METHODS I/R model was induced by bilateral carotid artery occlusion(BCAO)-reperfusion method and Y-maze learning and memory performance was assessed after reperfusion. Na^+-K^+-ATPase,Ca^(2+)-ATPase and SOD activity,as well as MDA content in mouse brain tissue were measured. Numbers of mouth-opening breaths of the isolated mouse head were observed in acute cerebral ischemia mice.RESULTS Learning and memory ability in mice with RCBE were improved significantly compared with model group. The activity of SOD,Na^+-K^+-ATPase and Ca^(2+)-ATPase were increased,while MDA contents decreased after RCBE(0.5,1.0 and 2.0 g·kg^(-1)) and piracetam(0.5 g·kg^(-1)) treatment compared with the model group. RCBE at all concentrations significantly prolonged the number of mouth-opening breaths of the isolated mouse head. CONCLUSION RCBE preconditioning exerts a marked neuroprotective effect on the ischemia brain,which is related to improve the learning and memory via regulating energy metabolism and anti-oxidation.

中草药徐长卿(Cynanchum paniculatum)治疗毒蛇咬伤的分子机制研究

徐长卿(Cynanchum paniculatum)作为一种传统中草药,被广泛应用于毒蛇咬伤治疗,然而其疗效的科学实验证据缺乏,治疗的分子机制也不清楚.本文采用有机溶剂萃取、柱层析及高效液相色谱法(HPLC)相结合,从徐长卿中筛选出和分离得到一个功效组分,经质谱和核磁共振鉴定为丹皮酚原苷.动物活体实验与组织病理切片观察表明,该组分显著地抑制尖吻蝮(Deinagkistrodon acutus)蛇毒引起的出血、水肿及肌肉组织坏死等活性.体外酶活检测显示,丹皮酚原苷可以抑制尖吻蝮蛇毒水解纤维蛋白原活性和PLA2活性.进一步采用分子模拟软件技术,揭示出丹皮酚原苷是通过与蛇毒中主要成分金属蛋白酶和PLA2发生相互作用,从而抑制尖吻蝮蛇毒发挥致伤作用.本研究首次揭示了中草药徐长卿治疗蛇伤的分子机制,为中医治疗毒蛇咬伤使用徐长卿找到了科学实验证据,也为蛇伤药物的研发提供了新的思路.

Recent advances in phytochemistry and pharmacology of C_(21) steroid constituents from Cynanchum plants

Cynanchum is one of the most important genera in Asclepiadaceae family, which has long been known for its therapeutic effects. In this genus, 16 species are of high medicinal value. The extracts of the root and/or rhizome parts have been applied in traditional Chinese medicines(TCM) for the prevention and treatment of various illnesses for centuries. C21 steroids, as the typical constituents of Cynanchum species, possess a variety of structures and pharmacological activities. This review summarizes the comprehensive information on phytochemistry and pharmacology of C21 steroid constituents from Cynanchum plants, based on reports published between 2007 and 2015. Our aim is to provide a rationale for their therapeutic application, and to discuss the future trends in research and development of these compounds. A total of 172 newly identified compounds are reviewed according to their structural classifications. Their in vitro and in vivo pharmacological studies are also reviewed and discussed, focusing on antitumor, antidepressant, antifungal, antitaging, Na+/K+-ATPase inhibitory, appetite suppressing and antiviral activities. Future research efforts should concentrate on in vitro and in vivo biological studies and structure activity relationship of various C21 steroid constituents.

Two new steroidal glycosides from the root of Cynanchum auriculatum

Two new steroidal glycosides named cyanoauriculoside A （1） and B （2）, have been isolated from the roots of Cynanchum auriculatum Royle ex Wight. The structure of cyanoauriculoside A （1） was determined to be 20-O-acetyl-penupogenin-3-O-β-D- glucopyranosyl-（ 1 →4）-β-D-glucopyranosyl-（ 1 →4）-α -L-cymaropyranosyl- （ 1 →4）-β-D-cymaropyranosyl- （ 1 →4）-α-L-diginopyranosyl-（1→4）-β-D-cymaropyranoside. Cyanoauriculoside B （2） was characterized as 20-O-acetylpenupogenin-3-O-β-D-gluco- pyranosyl-（ 1 →4）-α-L-cymaropyranosyl-（ 1 →4）-β-D-cymaropyranosyl-（ 1 →4）-α-L-diginopyranosyl-（ 1 →4）-β-D-cymaropyranoside. The structures of these compounds were elucidated based on the chemical and spectroscopic evidence.

Chemical constituents from endophytic fungus Plectosphaerella cucumerina YCTA2Z1 of Cynanchum auriculatum

Objective： To study the chemical constituents from the Et OAc extract of endophytic fungal Plectosphaerella cucumerina YCTA2 Z1.Methods： The chemical constituents were isolated and purified by silica gel column,Sephadex LH-20,and reverse-phase C-18 column chromatography as well as crystallization.Results： Thirteen compounds were isolated from the EtOAc extract of the fungal strain YCTA2 Z1.Their chemical structures were elucidated according to the spectral evidence.They were identified as caudatin（1）,baishouwubenzophenone（2）,cynandione B（3）,asterbatanoside A（4）,p-hydroxyphenethyl-O-β-D-glycoside（5）,caffeic acid（6）,ferulic acid（7）,2,5-dihydroxyacetophenone（8）,protocatechuic acid（9）,vanillic acid（10）,stearic acid（11）,azelaic acid（12）,and succinic acid（13）.Conclusions： It is the first chemical study on endophytic fungi from Cynanchum auriculatum and all the compounds are obtained from the species,the genus,as well as the family Plectosphaerellaceae for the first time.

A New Steroidal Glycoside from Roots of Cynanchum auriculatum

Objective To study the chemical constituents of Cynanchum auriculatum.Methods Two steroidal glycosides were purified by column chromatography and their structures were elucidated by spectroscopic analysis.Results and Conclusion Compound 1 is a new steroidal glycoside,named kidjoranin-3-O-β-D-cymaropyranoside.Compound 2 is isolated from this plant for the first time.

Application of a quantitative ^1H NMR method for rapid extraction and determination of the content of paeonol in Cynanchum paniculatum

Cynanchum paniculatum(Bunge) Kitagawa is usually used as an herbal medicine for treating many diseases. Paeonol is the main active component, and its content is the key indicator for quality control of C. paniculatum. In the present study, we developed a rapid, accurate and precise method for quantitation of paeonol in C. paniculatum using 1 H NMR spectra. The deuterated solvent of methanol-d4 enabled satisfactory separation of the signals to be integrated in 1 H NMR spectrum. H-6(δ 7.78) of 1 H NMR spectrum of C. paniculatum was selected as the feature signal for quantitation, and trimesic acid(TMA) was selected as an internal standard. Validation of the quantitative method was performed in terms of linearity, specificity, repeatability and stability. This is the first time to report quantitative 1 H NMR(qHNMR) applied to determine the content of paeonol in C. paniculatum and showed a wider linearity range than the reported quantitation of paeonol in others. The simple extraction of paeonol from C. paniculatum was rapid and will prompt the application of the developed method. This work implied that qHNMR represented a feasible alternative to HPLC-based methods for quantitation of paeonol in C. paniculatum, and it was suitable for the quality control of C. paniculatum.

Two new C_(21) steroidal glycosides isolated from Cynanchum komarovii

The present study was designed to further investigate the C_(21) steroidal glycosides in Cynanchum plants.Two new steroidal glycosides based on a 13,14:14,15-disecopregnane-type aglycone,komaroside P(1) and komaroside Q(2),together with three known compounds(3-5) were isolated from the whole herbs of Cynanchum komarovii.The aglycones of compounds 1 and 2 were two new disecopregnane.Their structures were elucidated on the basis of 1 D,2 D NMR spectroscopic data and acid hydrolysis.All the compounds(1-5) showed potent inhibitory activities against human leukemia cell lines(HL-60) with IC50 values ranging from 16.6 to 26.3 μmol·L^(-1),compared to the positive control 5-fluorouracil(6.4 μmol·L^(-1)).

Pharmacognostical study of Cynanchum stauntonii and Cynanchum glaucescens,botanical sources of TCM Baiqian

Objective： Cynanchum stauntonii and Cynanchum glaucescens are botanical species of Baiqian（Cynanchi Stauntonii Rhizoma et Radix） in Chinese Pharmacopoeia, in which, however, there are no microscopic identification. Therefore, we provided the morphological and microscopic identification of the crude drug for updating Chinese Pharmacopoeia.Methods： Twelve batches of C. stauntonii and three batches of C. glaucescens and their crude drugs were taxonomically, morphologically, and microscopically examined.Results： Taxonomically, C. stauntonii had narrowly lanceolate leaves with acuminate apex and 5 mm long petiole; Whereas C. glaucescens was oblong-lanceolate or oblong with rounded or acute apex in leaves,and had very short or no petiole. Morphologically, rhizomes of C. stauntonii and C. glaucescens both had hollow pith, but the hollow pith occupied about a half of the rhizome＇s diameter in C. stauntonii, whereas only a very small proportion of the overall diameter in C. glaucescens. Moreover, microscopic observation showed the difference in the proportion of xylem and in rhizome transverse-sections of the two species along with the difference in the size of the pith. Finally, laticifers and rhizome epidermal secretory cells were present in the powders of C. stauntonii, but absent from C. glaucescens.Conclusion： Based on observation of morphological and microscopic characteristics, the two species can be distinguished by the size of the pith, proportion of xylem of rhizomes, and crude drug powder characters such as laticifers and secretory cells.

Three New C_(21) Steroidal Glycosides from Roots of Cynanchum otophyllum

Objective To investigate the chemical structures of glycosides in the roots of Cynanchum otophyllum（Asclepiadaceae） and to find new glycosides. Methods The total glycosides in the roots of C. olophyllum were separated by silica gel column chromatography. The structures of the resulting compounds were determined by NMR and FAB-MS spectra. Results Three C21 steroidal glycosides were separated. Their structures were determined as caudatin 3-O-（4-O-methyl-β-D-cymaropyranosyl）-（1→4）-ɑ-D-oleandropyranosyl-（1→4）-β-D-glucopyranosyl-（1→4）-ɑ-L-rhamnopyranoside（1）, caudatin 3-O-β-D-digitoxopyranosyl-（1→4）-ɑ-D-oleandropyranosyl-（1→4）-β-Ddiginopyranosyl-（1→4）-β-D-glucopyranoside（2）, and caudatin 3-O-β-D-diginopyranosyl-（1→4）-ɑ-D-oleandropyranosyl-（1→4）-ɑ-D-oleandropyranosyl-（1→4）-β-Dglucopyranoside（3）, respectively. Conclusion Glycosides 1-3 are new compounds.