Synthesis, cytotoxicity, and hemolytic activity of 6'-O-substituted dioscin derivatives

Dioscin derivatives (1-12) with a variety of substitutions at the 6'-OH of the chacotriosyl residue and the 3',6'-anhydrosaponin derivatives (26, 30, and 32) were synthesized. All these derivatives showed much lower cytotoxicity than that of the parent dioscin, while their hemolytic activities were partially retained depending on the various 6'-O-substitutions.     

云南重楼ITS序列单核苷酸多态性与甾体皂苷特征的相关性分析

为了解云南重楼(Paris polyphylla var. yunnanensis) ITS序列的单核苷酸多态性(SNP)特征与其甾体皂苷构成特征的相关性,并探讨其对药材质量稳定性的影响,利用MegAlign软件对37份云南重楼样本的ITS序列进行对比,根据SNP位点进行分型;采用HPLC法测定7种甾体皂苷成分(重楼皂苷Ⅰ、Ⅱ、Ⅴ、Ⅵ、Ⅶ、H和薯蓣皂苷);用SPSS 25.0软件和SIMCA-P 15.0软件对各基因型甾体皂苷构成特征进行统计分析。结果表明,云南重楼ITS序列存在40个双等位多态性位点,其中碱基转换32个,碱基颠换8个,根据SNP特征可划分为2类基因型YN-I和YN-II。6种甾体皂苷(重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ、H、薯蓣皂苷)在云南重楼广泛分布,而重楼皂苷Ⅴ稀少。YN-I和YN-II的药典指标成分总含量分别为1.070%和0.93%,样本合格率分别为68.42%和77.78%;YN-I的甾体皂苷总含量为1.65%,YN-II为1.32%。方差分析表明,2类基因型的甾体皂苷特征存在一定程度的区别,但药典指标成分无显著差异,药材应该具有等效性。聚类分析和主成分分析表明,YN-Ⅰ的离散度更高,YN-Ⅱ的聚集度更好,表明YN-II的药材质量更加稳定,植株个体的甾体皂苷合成和累积差异更小,YN-II的SNP特征对云南重楼良种选育的分子遗传标记筛选具有重要意义。     

Protein-binding properties of a designed steroidal lactam compound

Introducing amide bonds into a steroid nucleus or its side chain may reduce the acute toxicity and enhance the pharmaceutical activity. In this work, a designed steroidal amide compound, named 3β-hydroxy-17-aza-d-homo-5-androsten-17-one (HAAO), was synthesized and identified. The interactions between HAAO and human serum albumin (HSA) were studied by multiple spectroscopic methods and molecular modeling procedures. It was found that HAAO locates in Sudlow’s site I in subdomain IIA of HSA molecules, relying on hydrogen bonds and van der Waals power to form HAAO–HSA complexes at ground state. The number of binding sites, binding constants, enthalpy change (ΔH^θ), Gibbs free energy change (ΔG^θ) and entropy change (ΔS^θ) were calculated at different temperatures based on fluorescence quenching theory and classical thermodynamic equation. The percentages content of the HSA’s secondary structures in presence of HAAO were detected by circular dichroism (CD) spectra and compared with those in no presence of HAAO. In addition, the experimental results of both binding site and conformational change were further confirmed by molecular modeling investigation, in which more details of the binding were visually unfolded. The information provided by the study may be useful for designing novel chemotherapeutic drugs and be helpful both in the early stages of drug discovery and in clinical practice.     

Microbial Baeyer–Villiger oxidation of 5α-steroids using Beauveria bassiana. A stereochemical requirement for the 11α-hydroxylation and the lactonization pathway

Beauveria bassiana KCH 1065, as was recently demonstrated, is unusual amongst fungal biocatalysts in that it converts C₁₉ 3-oxo-4-ene and 3β-hydroxy-5-ene as well as 3β-hydroxy-5α-saturated steroids to 11α-hydroxy ring-D lactones. The Baeyer–Villiger monooxygenase (BVMO) of this strain is distinguished from other enzymes catalyzing BVO of steroidal ketones by the fact that it oxidizes solely substrates with 11α-hydroxyl group. The current study using a series of 5α-saturated steroids (androsterone, 3α-androstanediol and androstanedione) has highlighted that a small change of the steroid structure can result in significant differences of the metabolic fate. It was found that the 3α-stereochemistry of hydroxyl group restricted “normal” binding orientation of the substrate within 11α-hydroxylase and, as a result, androsterone and 3α-androstanediol were converted into a mixture of 7β-, 11α- and 7α-hydroxy derivatives. Hydroxylation of androstanedione occurred only at the 11α-position, indicating that the 3-oxo group limits the alternative binding orientation of the substrate within the hydroxylase. Only androstanedione and 3α-androstanediol were metabolized to hydroxylactones. The study uniquely demonstrated preference for oxidation of equatorial (11α-, 7β-) hydroxyketones by BVMO from B. bassiana. The time course experiments suggested that the activity of 17β-HSD is a factor determining the amount of produced ring-D lactones. The obtained 11α-hydroxylactones underwent further transformations (oxy-red reactions) at C-3. During conversion of androstanedione, a minor dehydrogenation pathway was observed with generation of 11α,17β-dihydroxy-5α-androst-1-en-3-one. The introduction of C1C2 double bond has been recorded in B. bassiana for the first time.     

Urease inhibitory activity of ursane type sulfated saponins from the aerial parts of Zygophyllum fabago Linn

Five ursane type sulfated saponins have been isolated from the aerial parts of Zygophyllum fabago Linn. (locally called Chashum). The urease inhibitory effects of these compounds have been investigated for the first time as well as their molecular docking studies have also been carried out to check the structure–activity relationship. The IC₅₀ values of these compounds could not be found due to paucity of the samples. The molecular docking studies were performed only for the most active compound mono sodium salt of 3β,23-di-O-sulfonyl-23-hydroxyurs-20(21)-en-28-oic acid 28-O-[β-d-glucopyranosyl] ester (Zygofaboside A; 1).     

Natural products in Glycyrrhiza glabra (licorice) rhizome imaged at the cellular level by atmospheric pressure matrix‐assisted laser desorption/ionization tandem mass spectrometry imaging

The rhizome of Glycyrrhiza glabra (licorice) was analyzed by high‐resolution mass spectrometry imaging and tandem mass spectrometry imaging. An atmospheric pressure matrix‐assisted laser desorption/ionization imaging ion source was combined with an orbital trapping mass spectrometer in order to obtain high‐resolution imaging in mass and space. Sections of the rhizome were imaged with a spatial resolution of 10 μm in the positive ion mode, and a large number of secondary metabolites were localized and identified based on their accurate mass and MS/MS fragmentation patterns. Major tissue‐specific metabolites, including free flavonoids, flavonoid glycosides and saponins, were successfully detected and visualized in images, showing their distributions at the cellular level. The analytical power of the technique was tested in the imaging of two isobaric licorice saponins with a mass difference of only 0.02 Da. With a mass resolving power of 140 000 and a bin width of 5 ppm in the image processing, the two compounds were well resolved in full‐scan mode, and appeared with different distributions in the tissue sections. The identities of the compounds and their distributions were validated in a subsequent MS/MS imaging experiment, thereby confirming their identities and excluding possible analyte interference. The use of high spatial resolution, high mass resolution and tandem mass spectrometry in imaging experiments provides significant information about the biosynthetic pathway of flavonoids and saponins in legume species, combing the spatially resolved chemical information with morphological details at the microscopic level. Furthermore, the technique offers a scheme capable of high‐throughput profiling of metabolites in plant tissues.     

狭叶柴胡营养器官中柴胡皂苷和黄酮类化合物的积累部位及含量比较

采用植物解剖学、组织化学定位和紫外分光光度计分析方法,对狭叶柴胡各营养器官(根、茎、叶)的结构特征、营养器官中柴胡皂苷和黄酮类化合物的积累部位及其含量进行比较研究。结果表明:(1)狭叶柴胡根由周皮、中柱鞘薄壁组织和次生维管组织组成,茎由表皮、皮层和维管柱组成,叶为等面叶结构,其营养器官的结构均表现出对旱生环境的适应性。(2)组织化学定位显示,柴胡皂苷在狭叶柴胡根中主要分布在维管形成层和次生韧皮部及次生木质部靠近维管形成层的木薄壁细胞中,在茎中主要分布在表皮、皮层、维管形成层及韧皮薄壁细胞中,在叶中主要分布于表皮和叶肉中;狭叶柴胡的黄酮类化合物在茎中主要分布于表皮、棱角处的厚角组织及部分皮层细胞中,在叶中主要分布于表皮和位于叶缘及下表皮内的厚角组织中,在根内分布很少。(3)定量分析表明,柴胡总皂苷在狭叶柴胡根、茎、叶中的含量分别为2.635%、1.045%和0.981%;而黄酮类化合物在根、茎、叶中的含量分别为0.032%、1.212%和2.259%;定量分析结果与组织化学实验结果相符。研究表明,柴胡总皂苷在狭叶柴胡的根中含量最高,支持《中国药典》以根入药的结论;而且狭叶柴胡叶中的黄酮类化合物含量较高,建议对狭叶柴胡的叶进行综合开发利用。     

不同产地无患子果皮皂苷含量的地理变异研究

通过对不同产地无患子果皮皂苷含量的测定分析,结果表明,14个产地间果皮皂苷含量差异显著,平均含量为8.85%。果皮皂苷含量与产地经度间存在显著性正相关,东西部产地果皮皂苷含量差异明显,西部产地平均含量高于东部产地。聚类分析结果亦表明,东西部产地间果皮皂苷含量出现明显分化,西南部产地的江西宜丰、贵州榕江、广西龙州分化尤为明显,表现出较高的皂苷含量,为无患子高皂苷含量筛选的优良产区。     

药用植物甾体生物碱的药理作用及合成途径

甾体生物碱是药用植物中广泛存在的一类代谢产物,是一类具有降压、止咳、平喘、抗肿瘤等生物活性的天然产物。目前,甾体生物碱的合成代谢途径、分离纯化、鉴别及生物学功能研究已成为国内外天然产物研究的热点之一。对药用植物甾体生物碱的药理作用进行了综述,并根据萜类物质合成途径,推测总结了甾体生物碱的合成相关途径和参与该途径的关键酶及其基因克隆的研究进展,以期为药用植物甾体生物碱的代谢途径与基因表达调控及应用研究提供参考。     

Saponins in Tribulus terrestris – Chemistry and Bioactivity

Tribulus terrestris is a valuable herb known for its application in the folk medicine in many parts of the world. Furostanol and spirostanol saponins of tigogenin, neotigogenin, gitogenin, neogitogenin, hecogenin, neohecogenin, diosgenin, chlorogenin, ruscogenin and sarsasapogenin type are frequently found in this plant. Four sulphated saponins of tigogenin and diosgenin type are also isolated. Extracts and steroidal saponins have been found to possess various pharmacological activities. Preparations based on the saponin fraction of T. terrestris are used for treatment of infertility and libido disorders in men and women, as well as for treatment of cardiac diseases. Food supplements containing T. terrestris extracts are on sale in USA and Europe with claim of a general stimulating action. In this review the steroidal saponins found in T. terrestris are presented, covering the literature up to 2004. The data reveal clear difference in the saponin composition of samples from this plant species collected from different geographical regions. A comprehensive account on the biological activity of extracts and individual saponins is also included.