锉海绵属海绵Xestospongia sp.甾体类成分研究

目的 对广西涠洲岛北部湾海域来源的锉海绵属（Xestospongia）海绵中获得的结构新颖、活性良好的甾体类成分进行研究。方法 根据甾体类化合物的结构特性，利用多种柱色谱和高效液相色谱（HPLC）等方法，通过高分辨质谱（HRMS）、核磁共振（NMR）和旋光（ORD）等方法并结合文献对照对化合物进行结构鉴定。结果 从锉海绵属海绵中分离得到9个甾体化合物，分别为7-ketocholesterol (1)、3β-Hydroxy-24-methylene-5-cholesten-7-one (2)、(22E)-3β-Hydroxychoiesta-5,22-dien-7-one (3)、(22E)-3β-Hydroxychiest-5,22-diene-7,24-dione (4)、Dictyopterisin G (5)、Dictyopterisins D (6)、Dictyopterisins E (7)、antrocarine E (8)和Fucosterol (9)。结论 化合物2-8均为首次从该属海绵中分离得到，6和7母核上C-7位为少见的甲氧基，8是首次从海绵中分离得到5-OH和7羰基的甾体化合物。     

中国南海侧扁软柳珊瑚甾体类化学成分的研究

目的 研究侧扁软柳珊瑚（Subergorgia suberosa）的化学成分。方法 采用各种现代色谱技术对化学成分进行分离纯化,利用波谱技术鉴定其结构。结果 从侧扁软柳珊瑚的氯仿-甲醇提取物中分离并鉴定了8个甾体类化合物,分别为胆甾-5, 22-二烯-3β-醇（1）,麦角甾-5, 24(28)-二烯-3β-醇（2）,24-甲基胆甾-5, 22-二烯-3β-醇（3）,孕甾-3β-乙酰氧基-20-酮（4）,孕甾-1,4-二烯-3,20-二酮（5）,孕甾-6β-羟基-4-烯-3, 20-二酮（6）,孕甾-1,4-二烯-3-酮-20-羧酸甲酯（7）和9,11-开环甾醇3β, 6α, 11-trihydroxy-24-methyl-5α-cholest-9, 11-seco-7-en-9-one（8）。结论 所有化合物均为首次从该种珊瑚中分离得到。     

海绵共附生真菌Purpureocillium lilacinum的化学成分及T细胞调节活性研究

目的 对海绵Hymeniacidon sp.共附生真菌Purpureocillium lilacinum的化学成分及T细胞调节活性进行研究。方法 运用硅胶柱色谱、凝胶柱色谱及高压液相色谱等色谱技术对真菌乙酸乙酯粗提物进行分离纯化得到单体化合物,通过现代波谱（核磁共振,质谱）及光谱（旋光）的综合运用并结合文献对照对化合物结构进行鉴定,运用流式实验对化合物T细胞调节活性进行测试。结果 共分离获得5个甾体化合物,鉴定为(24R)-豆甾-4-烯-6β-醇-3-酮（1）、(22E,24R)-麦角甾-7,22-二烯-3β,5α,9α-三醇-6-酮（2）、(22E,24R)-麦角甾-7,22-二烯-3β,5α-二醇-6-酮（3）、麦角甾-5,24-二烯-3β-醇-7,23-二酮（4）及(22E,24R)-麦角甾-5,8,22-三烯-3β-醇-7-酮（5）。体外活性筛选实验中,化合物2~5在2.5 μmol/L时对刀豆蛋白A（ConA）诱导的T细胞增殖显示抑制活性,化合物5在10 μmol/L时对CD3+T细胞的增殖显示显著抑制活性,所有受试化合物对CD4+/CD8+ T细胞分化活性均无影响。结论 化合物1~4为首次从P. lilacinum中分离获得。本文是对化合物1~5 T细胞调节活性的首次报道。     

西瑁岛软珊瑚Sinularia sp. 的化学成分和生物活性研究

摘要：目的 对采自中国南海西瑁岛的软珊瑚Sinularia sp.的化学成分和生物活性进行研究。方法 综合利用薄层色谱、硅胶柱色谱、凝胶柱层析、半制备HPLC等分离手段,结合NMR和MS等波谱数据分析并对比文献数据,对化合物进行分离鉴定;选用A549和HL-60细胞株,采用CCK-8筛选方法,对化合物进行体外细胞毒活性测试。结果 从软珊瑚Sinularia sp.中共分离鉴定了10个甾体化合物,包括8个孕甾烷及其苷类化合物：3β-羟基-5,20-二烯孕甾烷乙酸酯（1）,20-烯-3-酮孕甾烷（2）,1,20-二烯-3-酮孕甾烷（3）,1,4,20-三烯-3-酮孕甾烷（4）,3β-羟基-20-烯孕甾烷（5）,3β-羟基-5,20-二烯孕甾烷（6）,1,4-二烯-3-酮-20α-羟基孕甾烷乙酸酯（7）,3β-羟基-5,20-二烯孕甾烷吡喃岩藻糖苷（8）;1个麦角甾烷类化合物：(22E,24R)-3β,5α,6β-三羟基-7,22-二烯麦角甾烷（9）和1个胆甾烷类化合物：(20S)-1,4-二烯-3,16-二酮-20β-羟基胆甾烷（10）。结论 化合物1～10对A549细胞株均未表现明显的细胞毒活性,但化合物4和7对HL-60细胞株具有中等的细胞毒活性,其IC50值分别为1.61 和 3.26 μmol/L。     

甾体衍生物的合成

目的:研究甾体甲基化转移酶。方法:从γ-oryzanol提取的cycloartenol和 24（28）-methylenecycloartenol为初始原料合成目标化合物。结果:成功地合成了11个甾体衍生物。结论:目标化合物均经HPLC 纯化;结构均经NMR和GC/MS确证,方法简便易行。     

云芝的化学成分

目的研究云芝的化学成分。方法用色谱学方法进行分离纯化,以波谱学方法进行结构解析。结果从云芝中分离并鉴定了八个化合物,分别为： 麦角甾-7,22-二烯-3β-棕榈酸酯（1）,麦角甾-7,22-二烯-3β-醇（2）,白桦脂酸（3）,4-羟基苯甲酸（4）,3-甲氧基-4-羟基苯甲酸（5）,3,5-二甲氧基-4-羟基苯甲酸（6）,2-呋喃酸（7）和烟酸（8）。结论八个化合物均为首次从云芝中分离得到。     

蒺藜果实中甾体皂苷类成分研究

目的研究蒺藜（Tribulus terrestrisL.）果实的皂苷类化合物。方法利用硅胶柱色谱、凝胶柱色谱、中低压色谱、高效液相色谱等手段进行分离,根据理化性质及波谱数据鉴定结构。结果从蒺藜果实中得到4个已知化合物,分别鉴定为（25R）-2α,3β-二醇-5α-螺甾烷-12-酮（门诺皂苷元）-3-O-{β-D-吡喃葡萄糖基-（1→2）-[β-D-吡喃木糖基-（1→3）]-β-D-吡喃葡萄糖基-（1→4）-β-D-吡喃半乳糖苷}（1）、26-O-β-D-吡喃葡萄糖基-（25R）-5α-呋甾-22-甲氧基-2α,3β,26-三醇-3-O-β-D-吡喃葡萄糖基-（1→2）-β-D-吡喃葡萄糖基-（1→4）-β-D-吡喃半乳糖苷（2）、海柯皂苷元-3-O-{β-D-吡喃葡萄糖基-（1→2）-[β-D-吡喃木糖基-（1→3）]-β-D-吡喃葡萄糖基-（1→4）-β-D-吡喃半乳糖苷}（3）、26-O-β-D-吡喃葡萄糖基-（25R）-5α-呋甾-12-羰基-20（22）-烯-3β,26-二醇-3-O-β-D-吡喃葡萄糖基-（1→4）-β-D-吡喃半乳糖苷（4）。结论化合物1和2为首次从该属植物中分离得到,化合物4为首次得到的25位R构型的甾体皂苷类单体化合物。     

百合中新的甾体皂苷的结构鉴定

百合中新的甾体皂苷的结构鉴定侯秀云１）陈发奎吴立军（沈阳药科大学中成药分析中药鉴定教研室，沈阳１１００１５）关键词百合；甾体皂苷百合为百合科植物百合（ＬｉｌｉｕｍｂｒｏｗｎｉＦ．Ｅ．Ｂｒｏｗｎｖａｒ．ｖｉｒｉｄｕｌｕｍＢａｋｅｒ．）的干燥肉质鳞片，具...     

穿龙薯蓣总皂苷中甾体皂苷的分离与鉴定

目的 研究穿龙薯蓣总皂苷中水溶性甾体皂苷，寻找新的活性化合物。方法 用硅胶柱色谱、薄层色谱及高效液相色谱等进行分离，通过酸水解、理化常数和波谱学分析（IR，NMR，MS，HMQC，HMBC）鉴定化合物结构。结果 从穿龙薯蓣总皂苷中分得2个甾体皂苷（1个水难溶性皂苷和1个水溶性皂苷），其化学结构分别鉴定为薯蓣皂苷元－3－O－{α-L-鼠李糖（1→2）－[β-D-葡萄糖（1→3）]}-β－D－葡萄糖皂苷（I），薯蓣皂苷元－3－O－[α－L－鼠李糖（1→3）－α-L-鼠李糖（1→4）－α－L－鼠李糖（1→4）]-β－D－葡萄糖皂苷（Ⅱ）。结论 Ⅱ为首次从穿龙薯蓣中分离得到的新化合物，命名为穿龙薯蓣皂苷Dc。     

微生物3-甾酮-△1-脱氢酶的研究进展

3-甾酮-△^1-脱氢酶（KSDH）属于黄素蛋白酶类，位于细胞膜上，能在3-甾酮化合物的C1和C2之间引入双键，提高原有底物的抗炎活性，在甾体药物的生产上发挥着重要作用。文章对已报道的不同微生物来源的KSDH进行比较分析，着重介绍了关于KSDH基因表达和敲除方面的研究，并对该酶在甾体转化方面的应用做一简要总结。     

Analysis of C-3 epimerization in (24R)-24,25-dihydroxyvitamin D3 catalyzed by hydroxysteroid dehydrogenase

Studies on the C-3 epimerization in (24R)-24,25-dihydroxyvitamin D(3) [24R,25(OH)(2)D(3)] were performed using hydroxysteroid dehydrogenases (HSDs). 3-Epi-24R,25(OH)(2)D(3) was formed from 24R,25(OH)(2)D(3) by the catalysis of 3alpha- or beta-HSD. These HSDs also catalyzed the C-3 epimerization in 3-epi-24R,25(OH)(2)D(3) to form 24R,25(OH)(2)D(3). 24R,25(OH)(2)D(3) and its C-3 epimer were separated by inclusion high-performance liquid chromatography using gamma-cyclodextrin (gamma-CD) as the mobile phase additive or a gamma-CD bonded chiral column. The product derived from the intermediate during the C-3 epimerization was isolated from the incubation specimens and identified as (7Z)-(24R)-24,25-dihydroxy-9,10-secocholesta-4,7,10(19)-trien-3-one by several instrumental analyses including (1)H-nuclear magnetic resonance spectrometry. The occurrence of this compound strongly proves that the formation of the C-3 epimer by HSD involves a dehydrogenation process. The present study suggests that HSDs may catalyze the C-3 epimerization of vitamin D compounds and modulate their concentrations and biological activities in animals and humans.     

Metabolism of 2 alpha-propoxy-1 alpha,25-dihydroxyvitamin D3 and 2 alpha-(3-hydroxypropoxy)-1 alpha,25-dihydroxyvitamin D3 by human CYP27A1 and CYP24A1.

Recently, we demonstrated that some A-ring-modified vitamin D3 analogs had unique biological activity. Of these analogs, 2alpha-propoxy-1alpha,25(OH)2D3 (C3O1) and 2alpha-(3-hydroxypropoxy)-1alpha,25(OH)2D3 (O2C3) were examined for metabolism by CYP27A1 and CYP24A1. Surprisingly, CYP27A1 catalyzed the conversion from C3O1 to O2C3, which has 3 times more affinity for vitamin D receptor than C3O1. Thus, the conversion from C3O1 to O2C3 by CYP27A1 is considered to be a metabolic activation process. Five metabolites were detected in the metabolism of C3O1 and O2C3 by human CYP24A1 including both C-23 and C-24 oxidation pathways. On the other hand, three metabolites of the C-24 oxidation pathway were detected in their metabolism by rat CYP24A1, indicating a species-based difference in the CYP24A1-dependent metabolism of C3O1 and O2C3 between humans and rats. Kinetic analysis revealed that the Km and kcat values of human CYP24A1 for O2C3 are, respectively, approximately 16 times more and 3 times less than those for 1alpha,25(OH)2D3. Thus, the catalytic efficiency, kcat/Km, of human CYP24A1 for O2C3 is only 2% of 1alpha,25(OH)2D3. These results and a high calcium effect of C3O1 and O2C3 in animal experiments using rats suggest that C3O1 and O2C3 are promising for clinical treatment of osteoporosis.     

Cimifoetisides VI and VII Two new cyclolanostanol triterpene glycosides from the aerial parts of Cimicifuga foetida

Two new cyclolanostanol triterpene glycosides, cimifoetiside VI (1) and cimifoetiside VII (2), and one known compound were isolated from the aerial parts of Cimicifuga foetida L. On the basis of spectral and chemical evidences, the structures of 1 and 2 were elucidated to be (23R,24S)-24-O-acetylisodahurinol-3-O-beta-d-galactopyranoside and (23R,24R)-24-O-acetylshengmanol-3-O-beta-d-glucopyranosyl-(1' --> 2')-beta-d-xylopyranoside. The known compound was identified as (23R,24R)-24-O-acetylshengmanol-3-O-beta-d-galactopyranoside (3).     

New cycloartane glycosides from Camptosorus sibiricus Rupr

Two new cycloartane glycosides were isolated from the whole herbs of Camptosorus sibiricus Rupr. By means of chemical and spectroscopic methods (IR, 1D and 2D NMR, HRMS, ESI MS), the structures were established as (24R)-3beta, 7beta, 24, 25-tetrahydroxycycloartane 3-O-beta-D-glucopyranosyl-24-O-beta-D-glucopyranoside (1) and (24R)-3beta, 7beta, 24, 25-tetrahydroxycycloartane 3-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-24-O-beta-D-glucopyranoside (2). At the same time, two new compounds were tested for their cytotoxicities in vitro against human tumor cell lines (A375-S2, Hela).     

Dammarane triterpenes from Ligustrum lucidum

Two new dammarane triterpenes 3beta-acetyl-20S,24R-dammarane-25-ene-24-hydroperoxy-20-ol (1), 20S,24R-dammarane-25-ene-24-hydroperoxy-3beta,20-diol (2), as well as three known dammarane triterpenes, 3beta-acetyl-20S,25-epoxydammarane-24alpha-ol (3), 20S,25-epoxydammarane-3beta,24alpha-diol (4), 20S-dammarane-23-ene-3beta,20,25-triol (5) were isolated from the fruits of Ligustrum lucidum. Compounds 3-5 were isolated from the fruits of L. lucidum for the first time. Their structures were elucidated by spectroscopic methods.     

Isolation and identification of 1alpha-hydroxy-24-oxovitamin D3 and 1alpha,23-dihydroxy-24-oxovitamin D3: metabolites of 1alpha,24(R)-dihydroxyvitamin D3 produced in rat kidney

1alpha,24(R)-Dihydroxyvitamin D3 [1alpha,24(R)(OH)2D3], a synthetic vitamin D3 analog, has been developed as a drug for topical use in the treatment of psoriasis. At present, the target tissue metabolism of 1alpha,24(R)(OH)2D3 is not understood completely. In our present study, we investigated the metabolism of 1alpha,24(R)(OH)2D3 in the isolated perfused rat kidney. The results indicated that 1alpha,24(R)(OH)2D3 is metabolized in rat kidney into several metabolites, of which 1alpha,24(R),25-trihydroxyvitamin D3, 1alpha,25-dihydroxy-24-oxovitamin D3, 1alpha,23(S),25-trihydroxy-24-oxovitamin D3, and 1alpha,23-dihydroxy-24,25,26,27-tetranorvitamin D3 are similar to the previously known metabolites of 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3]. In addition to these aforementioned metabolites, we also identified two new metabolites, namely 1alpha-hydroxy-24-oxovitamin D3 and 1alpha,23-dihydroxy-24-oxovitamin D3. The two new metabolites do not possess the C-25 hydroxyl group. Thus, the metabolism of 1alpha,24(R)(OH)2D3 into both 25-hydroxylated and non-25-hydroxylated metabolites suggests that 1alpha,24(R)(OH)2D3 is metabolized in the rat kidney through two pathways. The first pathway is initiated by C-25 hydroxylation and proceeds further via the C-24 oxidation pathway. The second pathway directly proceeds via the C-24 oxidation pathway without prior hydroxylation at the C-25 position. Furthermore, we demonstrated that rat kidney did not convert 1alpha-hydroxyvitamin D3 [1alpha(OH)D3] into 1alpha,25(OH)2D3. This finding indicates that the rat kidney does not possess the classical vitamin D3-25-hydroxylase (CYP27) activity. However, from our present study it is apparent that prior hydroxylation of 1alpha(OH)D3 at the C-24 position in the 'R' orientation allows 25-hydroxylation to occur. At present, the enzyme responsible for the C-25 hydroxylation of 1alpha,24(R)(OH)2D3 is unknown. Our observation that the side chain of 1alpha,24(R)(OH)2D3 underwent 24-ketonization and 23-hydroxylation even in the absence of the C-25 hydroxyl group suggests that 1alpha,25(OH)2D3-24-hydroxylase (CYP24) can perform some steps of the C-24 oxidation pathway without prior C-25 hydroxylation. Thus, we speculate that CYP24 may be playing a dual role in the metabolism of 1alpha,24(R)(OH)2D3.     

Apoptosis induced by uterine 24p3 protein in endometrial carcinoma cell line

The biological functions and reaction pathways of lipocalins in mammalian system were sought. Mouse uterine 24p3 protein is a secreted lipocalin from mouse uterus. To evaluate the effect of uterine 24p3 protein on the reproductive system, endometrial carcinoma cell line (RL95-2) was an experimental target for achieving the in vitro study. The cells were treated with 0.75 microM dexamethasone (DEX) or under serum-free medium to mimic the stress environment for various time periods, then employing Western blot to measure the 24p3 protein secretion. It showed the time-dependent induction effect on 24p3 protein and suggested the level of protein secretion correlating to environmental stress. Furthermore, the supplementation of 24p3 protein to the medium accompanied the reduction of cell viability. It showed that the 24p3 protein may be a death factor under conditional media via PI and annexinV-FITC assay. Based on the autocrine hypothesis, we investigated the effect of 24p3 protein on cultured RL95-2 cells upon the 24p3 protein interaction. We have demonstrated significant increase in intracellular reactive oxygen species upon 24p3 protein interaction. While the changes of mitochondrial membrane potential and cytochrome c release from mitochondria occurred, the activities of caspase-8, -9 and -3 were found to have increased. The condensation of DNA was occurred suggesting that 24p3 protein induced irreparable DNA damage, which in turn triggered the process of apoptosis. It shows evidence for the direct effect of this protein on endometrial cells. These findings suggest that 24p3 protein creates an intracellular oxidative environment that induces apoptosis in RL95-2 cells.     

Three new cycloartenol glycosides from the roots of Cimicifuga simplex

Three new cycloartenol triterpene saponins, 3 β,16 α-dihydroxy-12-acetoxy-16,22-cyclo-23-ketone-24 R,25-epoxy-cycloartane-7-ene 3- O- β-D-galactopyranoside ( 1), 24- O-hydroxy-7,8-didehydrohydroshengmanol 3- O- β-D-galactopyranoside ( 2), and 24-epi-24- O-hydroxy-7,8-didehydrohydroshengmanol 3- O- β-D-galactopyranoside ( 3), were isolated from the ethyl acetate soluble fraction of Cimicifuga simplex. Their structures were established by detailed spectroscopic analysis, including extensive 2D-NMR data. This is the first time that a 16,22-cyclo type glycosidesaponin from aCimicifuga species was reported. The immunosuppressive activities of the new compounds were evaluated by a ConA-stimulated T splenocyte proliferation assay in vitro.     

Bioactive steroids from the brown alga Sargassum carpophyllum

By activity-guided fractionation, two new sterols, 3beta,28xi-dihydroxy-24-ethylcholesta-5,23Z-dien (1) and 2a-oxa-2-oxo-5alpha-hydroxy-3,4-dinor-24-ethylcholesta-24(28)-ene (2), together with five known steroids, fucosterol (3), 24-ethylcholesta-4,24(28)-dien-3,6-dione (4), 24xi-hydroperoxy-24-vinylcholesterol (5), 24-ketocholesterol (6), 24R,28R- and 24S, 28S-epoxy-24-ethylcholesterol (7), were isolated from the brown alga Sargassum carpophyllum as active compounds causing morphological abnormality of Pyricularia oryzae mycelia. Compounds 1, 3, 4, 5 and 7 also exhibited cytotoxic activity against various cancer cell lines. The IC50 values for 1 and 5 against HL-60 were 7.8 and 8.5 microg/ml, 3 and 4 against P-388 were 0.7 and 0.8 microg/ml, whereas 7 against MCF-7, HCT-8, 1A9, HOS and PC-3 were 4.0, 8.8, 10.0, 10.0 and 7.2 microg/ml, respectively.     

New triterpenoid saponins from bulbs of Bolbostemma paniculatum

Nine new triterpenoid saponins were isolated from the bulbs of Bolbostemma paniculatum (Maxim.) Franquet (Cucurbitaceae): 7beta,18,20,26-tetrahydroxy-(20S)-dammar-24 E-en-3-O-alpha-L-(3-acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,18,20,26-tetrahydroxy-(20S)-dammar-24E-en-3-O-alpha-L-(4-acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,18,20,26-tetrahydroxy-(20S)-dammar-24E-en-3-O-alpha-L-arabinopyranosyl-(1-->2)-beta-D-(6-acetyl)glucopyranoside, 7beta,20,26-trihydroxy-(20S)-dammar-24E-en-3-O-alpha-L-arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,20,26-trihydroxy-(20S)-dammar-24E-en-3-O-alpha-L-(3-acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,20,26-trihydroxy-(20S)-dammar-24E-en-3-O-alpha-L-(4-acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,20,26-trihydroxy-8-formyl-(20S)-dammar-24E-en-3-O-alpha-L-(3-acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,20,26-trihydroxy-8-formyl-(20S)-dammar-24E-en-3- O-alpha-L-(4-acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside and 6'-O-palmitoyltubeimoside I. In addition, four known triterpenoid saponins: tubeimoside I, tubeimoside II, tubeimoside III and tubeimoside IV were isolated. The structures of the above compounds were elucidated based on spectroscopic studies, and the configuration of C-20 of tubeimoside IV was revised as S rather than R as reported in previous literature. The compounds were tested for their antiviral activity