黄皮种子中酰胺类生物碱及其杀线虫活性研究

为了解黄皮种子中的酰胺类生物碱及其杀线虫活性,运用多种色谱学及波谱学方法分离并鉴定了10个酰胺类生物碱,分别为:N-甲基桂皮酰胺(1),clausenalansamide A(2),3-dehydroxy-3-methoxyl-clausenalansamide A(3),clausenalansamide B(4),黄皮新肉桂酰胺B(5),N-(2-苯乙基)肉桂酰胺(6),2′-dehydroxy-2′-oxo-clausenalansamide B(7),lansamide-7(8),homoclausenamide(9),1,5-dihydro-5-hydroxy-1-methyl-3,5-diphenyl-2H-pyrrol-2-one(10)。其中,化合物3,7,10为新天然产物。首次对黄皮种子中的酰胺类生物碱2~8进行全齿复活线虫致死活性的测试,发现所测化合物均有致死活性,其中,化合物2,3,5和8有较强的致死活性,且均优于阳性对照除线磷,可为相关农药的研发提供科学依据。     

Novel steroidal saponins from Liriope graminifolia (Linn.) Baker with anti-tumor activities

Phytochemical investigation of the underground parts of Liriope graminifolia (Linn.) Baker resulted in the isolation of two new steroidal saponins lirigramosides A (1) and B (2) along with four known compounds. The structures were determined by extensive spectral analysis, including two-dimensional (2D) NMR spectroscopy and chemical methods, to be 3-O-{β-d-xylopyranosyl-(1→3)-α-l-arabinopyranosyl-(1→2)-[α-l-rhamnopyranosyl-(1→4)]-β-d-glucopyranosyl-(25S)-spirost-5-ene-3β,17α-diol (1), 1-O-[α-l-rhamnopyranosyl-(1→2)-β-d-xylopyranosyl]-(25R)-ruscogenin (2), 1-O-β-d-xylopyranosyl-3-O-α-l-rhamnopyranosyl-(25S)-ruscogenin (3), 3-O-α-l-rhamnopyranosyl-1-O-sulfo-(25S)-ruscogenin (4), methylophiopogonanone B (5), and 5,7-dihydroxy-3-(4-methoxybenzyl)-6-methyl-chroman-4-one, (ophiopogonanone B, 6), respectively. Compound 1 has a new (25S)-spirost-5-ene-3β,17α-diol ((25S)-pennogenin) aglycone moiety. The isolated compounds were evaluated for their cytotoxic activities against Hela and SMMC-7721 cells.     

New lanostane-type triterpenoids, inonotsutriols D, and E, from Inonotus obliquus

Two new lanostane-type triterpenoids, inonotsutriols D (1) and E (2), were isolated from the sclerotia of Inonotus obliquus (Pers.: Fr.) Pil. (Japanese name: kabanoanatake; Russian name: chaga). Their structures were determined to be lanost-8-ene-3β,22R,24R-triol (1) and lanost-8-ene-3β,22R,24S-triol (2) on the basis of spectral data, including 2D NMR analysis. In addition, major compounds, inotodiol (3), trametenolic acid (4), 3β-hydroxylanosta-8,24-dien-21-al (5), 21-hydroxylanosterol (6), inonotsuoxide A (7) and inonotsuoxide B (8) were identified, and all compounds, except 2, were evaluated for their cancer cell growth inhibitory activity against P388, HL-60, L1210 and KB cell lines.     

白花鬼针草中多烯炔类成分的分离及其生物活性研究

为了解白花鬼针草(Bidens pilosa var.radiata)的化学成分,采用多种色谱技术从其提取物中分离多烯炔类成分,并对其生物活性进行研究。结果表明,从白花鬼针草乙酸乙酯提取部位中分离鉴定出4个多烯炔类化合物,分别为5-acetoxy-2-phenylethinyl-thiophene (1)、1-phenylhepta-1,3,5-triyne (2)、5-phenyl-2-(1-propynyl)-thiophene (3)和icthyothereol acetate (4)。体外活性评价结果表明,化合物1～4均具有中等抗MRSA活性,且均对人肝LO_2细胞无毒性。首次为化合物1提供核磁数据并进行结构解析,化合物3、4为首次从该属植物中分离得到。     

稀有放线菌Lechevalieria rhizosphaerae NEAU-A2的次级代谢产物研究及遗传操作系统的建立

【背景】菌株Lechevalieria rhizosphaerae NEAU-A2是一株新的稀有放线菌,其基因组中包含多个与次级代谢产物生物合成相关的基因簇,具有产生丰富代谢产物的潜力。【目的】对稀有放线菌L. rhizosphaerae NEAU-A2的次级代谢产物进行研究,以期发现结构新颖或生物活性独特的化合物,并建立其遗传操作系统。【方法】应用硅胶柱色谱和高效液相色谱等方法对菌株NEAU-A2的次级代谢产物进行分离和纯化,整合质谱分析、核磁共振等方法进行结构解析。在对该菌株全基因组测序的基础上,以基因组序列中编码杂合的聚酮合酶-非核糖体肽合成酶(PolyketideSynthase-Nonribosomal Peptide Synthetase,PKS-NRPSs)基因1609为目标基因,利用PCR-Targeting介导的基因置换技术构建重组质粒,通过接合转移的方式导入野生菌株。【结果】从菌株NEAU-A2中分离鉴定了2个新的吲哚二聚化合物(1,2)和5个已知化合物N-乙酰色胺(3)、4-((2-(1H-Indol-3-Yl)Ethyl)Amino)-4-Oxobutanoic Acid (4)、Brevianamide F (5)、4S,7R-Germacra-1(10)E,5E-Diene-11-Ol (6)、1H-吡咯-2-羧酸(7)。接合子通过培养2代即可获得双交换突变菌株,PCR分析结果显示PKS-NRPS基因被成功中断。【结论】从稀有放线菌L. rhizosphaerae NEAU-A2中分离鉴定出7个化合物,并成功建立了该菌的遗传操作系统。     

Biotransformation of pseudolaric acid B by Chaetomium globosum

Pseudolaric acid B (1) is a natural product with potent antifungal activity. We discovered that pseudolaric acid B did not kill but only suppress the growth of the filamentous fungus Chaetomium globosum. It was proposed that pseudolaric acid B was converted to metabolites with decreased antifungal activities. In this study, a scaled-up biotransformation of pseudolaric acid B by C. globosum produced five metabolites, including three new compounds, pseudolaric acid I (2), pseudolaric acid B 18-oyl-alanine (4) and pseudolaric acid B 18-oyl-serine (6), together with two known compounds, pseudolaric acid F (3) and pseudolaric acid B 18-oyl-glycine (5). The structures were characterized by NMR and MS spectroscopy. The major biotransformation reaction was conjugation with amino acids. None of the metabolites showed inhibitory effects on the growth of Candida albicans. The results suggested that biotransformation might be a detoxification process for fungi to resist antifungal drugs.     

Biotransformation of gallic acid by <Emphasis Type="Italic">Beauveria sulfurescens</Emphasis> ATCC 7159

Preparative-scale fermentation of gallic acid (3,4,5-trihydroxybenzoic acid) (1) with Beauveria sulfurescens ATCC 7159 gave two new glucosidated compounds, 4-(3,4-dihydroxy-6-hydroxymethyl-5-methoxy-tetrahydro-pyran-2-yloxy)-3-hydroxy-5-methoxy-benzoic acid (4), 3-hydroxy-4,5-dimethoxy-benzoic acid 3,4-dihydroxy-6-hydroxymethyl-5-methoxy-tetrahydro-pyran-2-yl ester (7), along with four known compounds, 3-O-methylgallic acid (2), 4-O-methylgallic acid (3), 3,4-O-dimethylgallic acid (5), and 3,5-O-dimethylgallic acid (6). The new metabolite genistein 7-O-β-D-4″-O-methyl-glucopyranoside (8) was also obtained as a byproduct due to the use of soybean meal in the fermentation medium. The structural elucidation of the metabolites was based primarily on 1D-, 2D-NMR, and HRFABMS analyses. Among these compounds, 2, 3, and 5 are metabolites of gallic acid in mammals. This result demonstrated that microbial culture parallels mammalian metabolism; therefore, B. sulfurescens might be a useful tool for generating mammalian metabolites of related analogs of gallic acid (1) for complete structural identification and for further use in investigating pharmacological and toxicological properties in this series of compounds. In addition, a GRE (glucocorticoid response element)-mediated luciferase reporter gene assay was used to initially screen for the biological activity of the 6 compounds, 2–6 and 8, along with 1 and its chemical O-methylated derivatives 9–13. Among the 12 compounds tested, 11–13 were found to be significant, but less active than the reference compounds of methylprednisolone and dexamethasone.     

Limonoids from the fruits of Melia toosendan

Four (1–4) new and seven known limonoids were isolated from the EtOH extract of the fruits of Melia toosendan. The structures of the new compounds were established on the basis of spectroscopic methods to be 12-O-methyl-1-O-deacetylnimbolinin B (1), 12-O-methy-1-O-tigloyl-1-O-deacetylnimbolinin B (2), 12-O-ethylnimbolinin B (3), and 1-O-cinnamoyl-1-O-debenzoylohchinal (4). Additionally, two new tirucallane-type triterpenoids, named meliasenins S (5) and T (6), were obtained from the same fractions during purification of the limonoids.     

薏苡糠壳的化学成分及其种子萌发活性研究

为了解薏苡(Coixlachryma-jobi)糠壳的化学成分,利用多种柱色谱技术对其乙醇提取物乙酸乙酯萃取部位进行分离,经波谱数据分析鉴定了15个化合物,分别为香豆酸(1)、香豆酸甲酯(2)、2-羟乙基-香豆酸酯(3)、咖啡酸甲酯(4)、阿魏酸甲酯(5)、(E)-3-(4-甲氧基苯基)丙烯酸(6)、2,3-二羟基-1-(4-羟基-3-甲氧基苯基)-1-丙酮(7)、2,3-二羟基-1-(4-羟基-3,5-二甲氧基苯基)-1-丙酮(8)、对羟基苯甲酸(9)、3-羟基-4-甲氧基苯甲酸(10)、1,3,5-三甲氧基苯(11)、methyl (3-hydroxy-2-oxo-2,3-dihydroindol-3-yl)-acetate (12)、尿囊素(13)、2-(2-羟乙基)-3-甲基反丁烯二酸(14)和油酸(15),其中化合物3、7、12、13和14为首次从薏苡中分离得到。活性测试结果表明,化合物1、2、9、10和11对种子萌发具有较强的抑制作用。     

链霉菌CB02959中四霉素及四烯菌素的鉴定及培养基优化

【背景】四霉素(Tetramycin)和四烯菌素(Tetrin)是具有广谱抗真菌活性的四烯大环内酯类抗生素。链霉菌CB02959是一株雷纳霉素(Leinamycin)类化合物的潜在产生菌株,利用antiSMASH分析其基因组发现该菌株含有一个纳他霉素(Natamycin)类四烯大环内酯化合物的生物合成基因簇。【目的】对Streptomyces sp. CB02959中次级代谢产物进行研究,确定其是否可以产生四烯大环内酯化合物,对其发酵产物进行分离和结构鉴定,并进行初步的发酵优化以提高产量。【方法】基于生物信息学预测和高分辨质谱数据,推测CB02959中多烯化合物的结构;在不同发酵培养基中培养CB02959,确定适合大规模发酵的培养基;敲除tetrA基因以确定目标基因簇和四烯大环内酯化合物产生的相关性;分离和鉴定CB02959产生的主要代谢物的结构;通过改变培养基中葡萄糖、麦芽提取物和胰蛋白胨的含量,提高四烯大环内酯化合物的产量。【结果】通过对CB02959中纳他霉素类化合物生物合成基因簇的分析及16S rRNA基因序列的进化树分析,推测CB02959可能是一株新的四霉素和四烯菌素产生菌;在YEME发酵培养基中对CB02959进行大规模发酵,分离得到4个化合物,鉴定为四霉素A (1)、四霉素B (2)、四烯菌素A (3)、四烯菌素B (4);最后通过培养基的初步优化,将化合物1–4的产量分别提高至208.1、100.0、1 315.6、109.9 mg/L。【结论】通过基因组挖掘策略发现了一株新的四霉素和四烯菌素产生菌链霉菌CB02959,并通过培养基优化提升了其四烯大环内酯化合物的产量,此发现为这类抗真菌天然产物的后续开发奠定了基础。     

铁皮石斛内生真菌次生代谢产物研究

为了解铁皮石斛(Dendrobium officinale)内生真菌Phyllosticta aristolochiicola的次生代谢产物,从该真菌中分离得到15个化合物,经波谱分析分别鉴定为N-methyl-2-pyrolidinone (1)、环-(甘氨酸-L-脯氨酸)(2)、环-(D-丙氨酸-L-脯氨酸)(3)、环-(L-缬氨酸-L-脯氨酸)(4)、环-(L-亮氨酸-L-脯氨酸)(5)、cyclo-(L-Leu-D-4-hydroxyprolinyl)(6)、环-(L-苯丙氨酸-L-脯氨酸)(7)、环-(L-苯丙氨酸-L-4-羟基脯氨酸)(8)、环-(L-酪氨酸-L-脯氨酸)(9)、环-(L-苯丙氨酸-L-亮氨酸)(10)、啤酒甾醇(11)、对羟基苯乙醇(12)、对羟基苯乙酸(13)、(2S,3R)-1-(4-羟基苯基)丁烷-2,3-二醇(14)和(2R,3S)-1-苯基丁烷-2,3-二醇(15)。采用MTS法检测抗肿瘤活性表明,化合物2、10和14对HL-60、A-549、SMMC-7721、MCF-7和SW-480细胞株具有一定的抑制活性。     

Alkyl phenols and saponins from the roots of Labisia pumila (Kacip Fatimah)

Recently, there is a remarkable boom in the sales of Labisia pumila (Kacip Fatimah) in the Malaysian market, as an extract of the plant is used to gain energy and libido as well as to treat many other ailments. A chemical analysis of its roots was undertaken and three metabolites, demethylbelamcandaquinone B (1), fatimahol (2), and dexyloprimulanin (3) together with 21 known compounds including epoxyoleanane glycosides, alkenated phenolics, cerebroside, glycerogalactolipids, and lipids were isolated and identified. Structure elucidation was achieved by spectroscopic and chemical studies. The MeOH extract of KF and compounds 12 and 13 exhibited moderate in vitro antibacterial activity.     

柯拉斯那沉香的生物活性成分研究

为了解柯拉斯那(Aquilaria crassna)的化学成分,从其所产沉香中分离得到10个化合物,经波谱分析分别鉴定为:6,8-羟基-2-(2-苯乙基)色酮(1),6,8-二羟基-2-[2-(4-甲氧基苯)乙基]色酮(2),rel-(1a R,2R,3R,7b S)-1a,2,3,7b-tetrahydro-2,3-dihydroxy-5-(2-phenylethyl)-7H-oxireno[f][1]benzopyran-7-one(3),rel-(1a R,2R,3R,7b S)-1a,2,3,7b-tetrahydro-2,3-dihydroxy-[2-(4-methoxyphenyl)-ethyl]-7H-oxireno[f][1]benzopyran-7-one(4),rel-(1a R,2R,3R,7b S)-1a,2,3,7b-tetrahydro-2,3-dihydroxy-5-[2-(3-hydroxy-4-methoxyphenyl)-ethyl]-7H-oxireno[f][1]benzopyran-7-one(5),oxidoagarochromone B(6),oxidoagarochromone C(7),(5S,6R,7S,8R)-2-[2-(3′-hydroxy-4′-methoxyphenyl)ethyl]-5,6,7,8-tetrahydroxy-5,6,7,8-tetrahydrochromone(8),6,7-cis-dihydroxy-2-(2-phenylethyl)-5,6,7,8-tetrahydrochromone(9),N-trans-feruloyltyramine(10)。化合物3~5和8~10为首次从柯拉斯那沉香中分离得到。化合物1,3,6,7,9和10对乙酰胆碱酯酶具有一定的抑制活性,化合物4对人慢性髓原白血病细胞株K-562和人胃癌细胞株SGC-7901均具有较小的抑制作用,化合物1和3对人肝癌细胞株BEL-7402也有抑制活性。     

苦槛蓝叶中的黄酮类化合物

为了解苦槛蓝(Myoporum bontioides A.Gray)的化学成分,采用色谱分离技术从苦槛蓝叶片中分离得到11个黄酮类化合物。通过波谱分析,他们的结构分别鉴定为:桔皮素(1)、甜橙素(2)、5,4′-二羟基-6,7,8,3′-四甲氧基黄酮(3)、4′,5,7,8-四甲氧基黄酮(4)、去甲基川陈皮素(5)、5-羟基-6,7,3′,4′-四甲氧基黄酮(6)、3′,4′,5,6,7,8-六甲氧基黄酮(7)、二氢山柰酚(8)、木犀草素(9)、3′,4′,5,7-四羟基-3-甲氧基黄酮(10)和芹黄素(11)。除化合物9之外,其他化合物均为首次从苦槛蓝叶片中分离得到。孢子萌发法测定结果表明,化合物1,2,8和9对香蕉炭疽菌(Colletotrichum musae)具有较好的抑菌活性。这为苦槛蓝叶片中有效成分的利用提供了理论依据。     

Hybrid flavan-chalcones, aromatase and lipoxygenase inhibitors, from Desmos cochinchinensis

Hybrid flavan-chalcones, desmosflavans A (1) and B (2), together with three known compounds, cardamonin (3), pinocembrin (4) and chrysin (5), were isolated from leaves of Desmos cochinchinensis. Cardamonin (3) and chrysin (5) exhibited potent antioxidant activity with 15.0 and 12.2 ORAC units. Desmosflavans A (1) and B (2), pinocembrin (4), and chrysin (5) were found to be inhibitors of aromatase with respective IC₅₀ values of 1.8, 3.3, 0.9, and 0.8 μM. Desmosflavan A (1) inhibited lipoxygenase with the IC₅₀ value of 4.4 μM. Desmosflavan A (1) exhibited cytotoxic activity with IC₅₀ values of 0.29–3.75 μg/mL, while desmosflavan B (2) showed IC₅₀ values of 1.71–27.0 μg/mL.     

毛果鱼藤化学成分及生物活性研究

为了解毛果鱼藤(Derris eriocarpa How)藤茎和根中的生物活性成分,采用柱色谱技术,从其乙酸乙酯萃取部位分离得到6个化合物,分别鉴定为:高丽槐素(1)、美迪紫檀素(2)、大黄素(3)、松脂醇(4)、(6R,9R)9-hydroxy-4-megastigmen-3-one(5)、2-methoxygliricidol(6)。这些化合物均为首次从该植物中分离得到。体外抑菌、细胞毒抑制活性测试结果表明,化合物1、2对部分肿瘤细胞株及结核分枝杆菌(H37Rv)有明显抑制活性。     

喙果皂帽花根的化学成分研究

为了解喙果皂帽花(Dasymaschalon rostratum)根的化学成分,采用多种色谱技术从其根的乙醇提取物中分离得到8个化合物。通过波谱数据分析,分别鉴定为:花旗松素(1)、(+)-儿茶素(2)、毛蕊异黄酮(3)、(-)-丁香树脂酚(4)、(-)-皮树脂醇(5)、lirioresinol A(6)、N-反式肉桂酸酰对羟基苯乙胺(7)和羟基酪醇(8)。所有化合物均为首次从该植物中获得。化合物1~8均无明显抗菌活性。     

华石斛化学成分研究

为了解华石斛(Dendrobium sinense)的化学成分,采用多种柱色谱技术从其全草乙醇提取液中分离纯化了10个化合物,经波谱分析分别鉴定为:鼓槌石斛素(1)、2′,4′-二羟基查尔酮(2)、2,5,7-三羟基-4-甲氧基-9,10-二氢菲(3)、4,7-二羟基-2,3-二甲氧基-9,10-二氢菲(4)、2,5-二羟基-3,4-二甲氧基-9,10-二氢菲(5)、2,7-二羟基-3,4,6-三甲氧基-9,10-二氢菲(6)、(E)松柏醛(7)、反式对羟基肉桂酸酯(8)、对羟基苯丙酸甲酯(9)和十二元内环酯(10)。所有化合物均为首次从华石斛中分离得到,其中化合物2、6、7和10对乙酰胆碱酯酶具有一定的抑制活性。     

喙尾琵琶甲肠道来源链霉菌(Streptomyces sp.)BPA71次级代谢产物的分离鉴定及其生物活性评价

【背景】昆虫是世界上种类最多、肠道菌群资源最丰富且多样的动物类群之一。昆虫肠道微生物具有产生活性次级代谢产物的能力,是活性天然产物的重要来源。【目的】研究药用昆虫喙尾琵琶甲(Blaps rynchopetera)成虫肠道来源链霉菌(Streptomyces sp.) BPA71的次级代谢产物及其生物活性。【方法】利用正相硅胶柱色谱、葡聚糖凝胶Sephadex LH-20柱色谱等方法分离纯化该菌株的发酵粗提物,采用牛津杯法进行抗菌活性追踪,确定抗菌活性部位,通过ESI-MS、NMR等波谱数据分析对化合物结构进行鉴定,采用微量肉汤稀释法测定最低抑菌浓度(minimal inhibitory concentration, MIC),采用MTS法测定抗肿瘤活性。【结果】从Streptomyces sp. BPA71的固体发酵提取物中共分离得到4个已知化合物,通过对比核磁数据确定为糠酸甲酯(1)、吡咯甲酰胺A (2)、吡咯甲酰胺B (3)和吲哚-3-乙酸甲酯(4)。抗菌活性结果显示化合物2具有广谱抗菌活性。此外,化合物2对宫颈癌细胞HeLa、肺癌细胞A549、肝癌细胞SMMC-7721、乳腺癌细胞MDA-MB-231和结肠癌细胞SW480这5株肿瘤细胞均有明显的抑制活性。【结论】喙尾琵琶甲肠道来源Streptomyces sp. BPA71可产生丰富的生物活性物质,该研究结果为进一步挖掘喙尾琵琶甲肠道链霉菌的活性天然产物奠定了基础,同时丰富了人们对喙尾琵琶甲肠道微生物的认识。     

人工诱导海南龙血竭的化学成分研究

为了解人工诱导海南龙血树(Dracaena cambodiana)所产血竭的化学成分,从其乙醇提取物中分离得到10个化合物,经波谱分析分别鉴定为socotrin-4?-ol(1)、homoisosocotrin-4?-ol(2)、(E)-3-(3,4-dihydroxybenzylidene)-7-hydroxy-chroman-4-one(3)、5-hydroxy-7-methoxy-3-(4?-hydroxybenzyl)-4-chromanone (4)、3-去氧苏木查耳酮(5)、苏木查耳酮(6)、7,4?-二羟基黄酮(7)、7,4?-二羟基-8-甲基黄酮(8)、丁香树脂醇(9)和邻苯二甲酸二(2-乙基己基)酯(10)。化合物1～10均为首次从人工诱导海南龙血树所产血竭中分离得到,其中化合物8为新天然产物,化合物3～6为首次从血竭中分离得到。化合物7和8对耐甲氧西林金黄色葡萄球菌具有生长抑制作用。