青霉属真菌Penicillium sp. CPCC 400786的抗病毒活性成分

采用抗艾滋病毒抑制剂筛选模型对一株青霉属真菌Penicillium sp. CPCC 400786发酵产物的乙酸乙酯提取物进行活性评价,结果显示,其对艾滋病毒有较强的抑制活性。采用正相硅胶柱、Sephadex LH-20凝胶柱和半制备HPLC等色谱技术对乙酸乙酯提取物进行分离纯化,从中分离得到8个化合物。通过波谱数据分析,分别鉴定为：oxalicine A（1）、oxalicine B（2）、cis-4,6-dihydroxymellein（3）、亚油酸（4）、十八烯酸（5）、肉豆蔻酸（6）、尿嘧啶（7）、胸腺嘧啶（8）。化合物1和2为杂萜类化合物。对化合物1-6进行了抗艾滋病毒（HIV-1）和抗甲型流感病毒（H1N1）的活性评价。结果显示,化合物1具有良好的抗H1N1活性,其IC₅₀值为38.5μmol/L,比阳性对照药利巴韦林稍弱（IC₅₀=20.5μmol/L）;化合物1和2具有抗HIV-1的活性,其IC₅₀值分别为22.4、67.8μmol/L;其他化合物未显示抗病毒活性。本研究为从青霉属中发现更多抗病毒活性杂萜分子提供了依据。     

海洋真菌Aspergillus niger XJJ-3中萘并吡喃酮类化合物结构及生物活性

【背景】萘并吡喃酮类化合物生物活性多样,是真菌Aspergillusniger中特征次生代谢产物。【目的】研究分离自海洋滩涂土壤的真菌Aspergillus niger XJJ-3中萘并吡喃酮类化合物结构及其抗菌和卤虫致死活性。【方法】以TLC分析为导向,综合运用多种色谱和光谱方法分离和鉴定萘并吡喃酮类化合物。采用微量稀释法测试化合物的抗菌和卤虫致死活性。【结果】从真菌A.niger大米发酵产物中共分离得到6个萘并吡喃酮类化合物,鉴定为RubrofusarinB(1)、Flavasperone (2)、Aurasperone A (3)、Asperpyrones C (4)、Asperpyrones B (5)和Fonsecinone A (6)。抗菌活性实验结果表明,化合物1-6对致病菌S. aureus ATCC33591、29213、E. faecium ATCC35667和V.parahemolyticus表现出不同程度的抑制活性,其中化合物2和4对S.aureus ATCC33591表现出较强抑制活性(MIC分别为43.7μmol/L和21.9μmol/L),化合物3对E.faecium ATCC35667抑制活性较强(MIC为21.9μmol/L)。卤虫致死活性实验结果表明,化合物1-6均表现出一定的卤虫致死活性,其中化合物2和3活性显著(LD50分别为35.0μmol/L和8.8μmol/L)。【结论】菌株XJJ-3可产生结构丰富的萘并吡喃酮类化合物,化合物1-6存在不同程度的抗菌和卤虫致死活性,该研究可为抗菌和细胞毒类药物的研发提供参考。     

操纵茶树类黄酮3′-羟基化酶生物合成B环-3′,4′-二羟基黄酮类化合物

【目的】操纵茶树类黄酮3′-羟基化酶,生物合成B环-3′,4′-二羟基黄酮类化合物圣草酚、二氢槲皮素和槲皮素。【方法】构建了4个茶树类黄酮3′-羟基化酶基因(CsF3′H)和拟南芥的P450还原酶基因(ATR)融合表达质粒:SUMO-CsF3'H[7-517]::ATR1[49-688]3 AA、SUMO-CsF3'H[28-517]::ATR1[49-688]3 AA、SUMO-CsF3'H[7-517]::ATR2[75-711]3 AA和SUMO-CsF3'H[28-517]::ATR2[75-711]3 AA,分别转化大肠杆菌菌株TOP10、DH5α和BL21,获得12个转化菌株S1–S12;构建了茶树类黄酮3′-羟基化酶基因CsF3′H表达质粒p YES-Dest52-CsF3′H,转化酵母菌株WAT11,得到转化菌株S13;构建了茶树类黄酮3′-羟基化酶基因CsF3′H表达质粒pES-URA-CsF3′H,及茶树黄烷酮3-羟基化酶基因CsF3H与拟南芥黄酮醇合成酶基因At FLS的融合表达质粒pES-HIS-CsF3H::At FLS 9AA,二者共转化酵母菌株WAT11,获得转化菌株S14。【结果】转化SUMO-CsF3'H[28-517]::ATR1[49-688]3 AA质粒的TOP10菌株S6在25°C条件下发酵,转化效率最高,能将1000μmol/L柚皮素、二氢山奈酚和山奈酚,分别转化生成287.93μmol/L圣草酚、131.76μmol/L二氢槲皮素和188.62μmol/L槲皮素。发酵菌株S13能分别将1000μmol/L柚皮素、二氢山奈酚和山奈酚,最多能转化生成734.32μmol/L圣草酚、446.07μmol/L二氢槲皮素和594.64μmol/L槲皮素。喂食S14发酵菌株5 mmol/L的底物柚皮素,在发酵36–48 h中,最多能生成1412.16μmol/L圣草酚、490.25μmol/L山奈酚、445.75μmol/L槲皮素、66.75μmol/L二氢槲皮素和73.50μmol/L二氢山奈酚。【结论】本研究首次将茶树类黄酮3′-羟基化酶基因应用于B环-3′,4′-二羟基黄酮类化合物圣草酚、二氢槲皮素和槲皮素的生物合成。     

一株毛壳霉属真菌中新结构活性聚酮类化合物研究

采用硅胶、凝胶柱层析以及反相高效液相色谱等分离技术,从一株毛壳霉属真菌的固体发酵提取物中分离纯化了5个聚酮类化合物chaetomones A-E(1-5),其结构主要通过分析核磁共振数据确定。生物活性测试结果表明化合物5具有抗白色念珠菌Candida albicans(ATCC10231)的活性,其IC50为20.0μmol/L。     

小麦幼苗及不同育性花药中P5C还原酶的研究

从小麦幼苗和花药中提取的二氢吡咯-5-羧酸(P5C)还原酶,在可育花药中活性很高,约为幼苗中活性的7～13倍,表明花药有很高的脯氨酸合成能力。从减数分裂期到单核靠边期酶活性逐渐升高,到双核初期明显降低。在不育花药中,减数分裂期酶活性高于可育花药,到单核初期酶活明显下降,仅为可育花药活性的一半。 初步纯化的小麦幼苗P5C还原酶的最适pH为7.2左右。对P5C和NADH的K_m值分别为400μmol/L和370/μmol/L。以NADPH为供氢体,其酶活性为NADH的35％。NADP~+、NAD~+、ATP、ADP对酶活性均有强烈的抑制作用。脯氨酸浓度在10m mol/L以上对酶活有轻微抑制作用。     

白桦树羽扇豆烷型三萜类化合物与抑制PTP1B活性研究

蛋白酪氨酸磷酸酯酶1B(Protein tyrosine phosphatase1B;PTP1B)在胰岛素信号传递过程中起负调控作用,亦是研究治疗2型糖尿病的重要靶点。利用PTP1B生物活性导向分离方法,从白桦树皮的正己烷提取物中分离得到7个羽扇豆烷型三萜类化合物。经过光谱分析和文献比较,确定分离得到的7个化合物为:羽扇烯酮(1),羽扇豆醇(2),桦木酸(3),白桦脂醛(4),白桦脂酸(5),算盘子酮醇(6)和白桦脂醇(7)。其有效抑制PTP1B活性值(IC50)分别为:5.6±0.3μmol/L,4.1±0.2μmol/L,7.2±0.3μmol/L,12.6±0.4μmol/L,11.6±0.3μmol/L,9.6±0.4μmol/L和13.6±0.5μmol/L。     

铁和镍对光合细菌生长和产氢的影响

基于金属元素在生物体功能发挥中的作用以及它们参与光合细菌光合放氢的重要性,着重进行了铁和镍对沼泽红假单胞菌（Rhodopseudomonas palustris）Z菌株和一株红杆菌（Rhodobactersp.）细胞生长、光合放氢和光合色素合成影响的研究。结果表明,高浓度Fe3+可显著提高两菌株光放氢能力和生物合成能力,最适浓度的Fe3+可使其产氢能力分别达对照组的1.32倍和2.8倍,产氢得率分别为360.6mL/g和385.9 mL/g,生物量分别为对照组的1.42倍和1.54倍。9μmol/L Ni2+的添加可使两菌株产氢能力分别达对照组的1.48倍和1.96倍,产氢得率分别为429.7mL/g和456.3 mL/g。而当Ni2+浓度为12μmol/L时,两菌株的产氢活性受到不同程度的抑制,产氢得率分别降低46.7%和19.4%。在铁浓度相同时,添加6μmol/L Ni2+能明显促进两菌株的生长。而当Ni2+浓度大于6μmol/L时,细胞生长受到抑制。Fe3+和Ni2+对Rhodobactersp.菌株类胡萝卜色素有显著影响。研究结果显示, 426nm色素峰随铁浓度的增加和镍的添加而消失,同时,产氢活性提高。     

海洋真菌杂色曲霉F62丁内酯类化合物研究

为了深入研究相似蜂海绵相关真菌杂色曲霉F62的活性代谢产物,采用硅胶柱、凝胶柱色谱和HPLC等分离手段对其大米固体发酵物的乙酸乙酯提取物进行分离,并利用质谱和核磁共振等现代波谱学技术对其结构进行鉴定,从中共得到6个丁内酯类化合物,分别为丁内酯Ⅰ(1)、丁内酯Ⅱ(2)、丁内酯Ⅲ(3)、丁内酯Ⅳ(4)、丁内酯Ⅶ(5)、Aspernolides A(6),其中化合物1在10μmol/L时表现出较强的抗炎活性,其IC50值为8.73μmol/L。     

3β,20α-羟基甾体脱氢酶的动力学性质

3β,20α-羟基甾体脱氢酶(3β,20α-Hydroxysteroid dehydrogenase,3β,20α-HSD)是从胎羊血中分离得到的。分子量为35kD。该酶以NADPH为辅酶,有两种底物。以孕酮为底物时,Km=30.8μmol/L,Vmax=0.7nmol min~(-1)(nmol enzyme)~(-1);以5α-二氢睾酮(5α-Dihydrotestosterone,5α-DHT)为底物时,Km=74μmol/L,Vmax=1.3nmol min~(-1)(nmol enzyme)~(-1)。5α-DHT竞争性抑制20α-还原活性,Ki=102μmol/L。16α-溴代乙酰氧基(16α-Bromo acetoxyprogesterone,16α-BAP)是3β,20α-HSD不可逆竞争性抑制剂,t_(1/2)=75min。对3β和20α还原活性的抑制常数Ki分别为23μmol/L和58μmol/L。     

三白草中ATG4B抑制剂的发现及活性测定

验证从三白草中提取的两个化合物XGN56和XGN59对自噬关键蛋白ATG4B酶活性的影响及对自噬的调节作用。分子对接的方法验证化合物与游离ATG4B及ATG4B-LC3复合体的氢键结合作用;SDS-PAGE法及荧光共振能量转移法(FRET)测定化合物(10μmol/L)抑制ATG4B的IC50值;LC3融合GFP荧光标签检测化合物(10μmol/L)对LC3荧光聚集的影响,并设置正常组、给药组和药物联用Baf(0.5μmol/L)组;过表达GFP-LC3的WT-MEF及ATG5-/--MEF细胞检测化合物诱导LC3荧光点的情况。结果显示,XGN56和XGN59能分别与游离ATG4B和ATG4B-LC3复合体形成氢键作用,且两者均能剂量依赖地抑制ATG4B的酶切活性,体外IC50分别为7.74μmol/L和8.00μmol/L,同时能够ATG5依赖地促进GFP标记的自噬体的生成(P<0.001)。结果表明,两个化合物可能是通过一定程度地抑制ATG4B的酶活性从而促进细胞自噬水平。     

Synthesis,anti-diabetic evaluation and molecular docking studies of 4-(1-aryl-1H-1, 2, 3-triazol-4-yl)-1,4-dihydropyridine derivatives as novel 11-β hydroxysteroid dehydrogenase-1 (11β-HSD1) inhibitors

11-Beta-Hydroxysteroid dehydrogenase-1(11β-HSD1) inhibitors are one of the emerging classes of molecules to fight against diabetic complications. A novel series of 4-(1-substituted-1H-1,2,3-triazol-4-yl)-1,4-dihydropyridine derivatives were synthesized and evaluated for their anti-diabetic activity. Two compounds showed anti-diabetic activity very effectively. To clarify the mechanism of action of these compounds, the most potent compounds (5g and 5h) of the synthesized analogs were further studied by testing its 11-Beta Hydroxysteroid dehydrogenase-1 inhibitory activity through in vitro enzymatic experiments. The results showed that the 11β-HSD1 inhibitory activity of compounds 5g and 5h was stable and efficient. Molecular docking studies revealed compounds 5g (−9.758) and 5h (−8.495) to have a stable binding patterns to the human 11-Beta-Hydroxysteroid dehydrogenase-1.     

3S-(+)-3,7-Dimethyl-1,5-Octadiene-3,7-Diol and Ionone Derivatives from Tea

1-Isobutyl-5-(4-phenoxyphenyl)imidazole (KK-98), an inhibitor of juvenile hormone (JH) biosynthesis in the cockroach, and related imidazole compounds were evaluated against silkworm, Bombyx mori, for their activity to induce precocious metamorphosis. KK-98 induced precocious metamorphosis in the 4th instar larvae at high doses. Replacement of the 4-phenoxy group by a 3-phenoxy or 3-benzyloxy group on the benzene ring increased the activity. Among this series of compounds, 5-(3-benzyloxyphenyl)-1-isopropylimidazole (8) showed the highest activity. The induction of precocious metamorphosis by compound 8 was rescued by the simultaneous application of methoprene, a JH minie. When newly molted 3rd instar larvae were treated with a high dose of compound 8, a few larvae formed larval-pupal intermediates in the 3rd instar stage, which has not been formed by treating of any other imidazoles so far.     

Mutagenic activity of carcinogenic and noncarcinogenic nitrofurans and of urine of rats fed these compounds

The nitrofurans, 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2), N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT), nitrofurantoin, 5-nitro-2-furoic acid, 5-nitro-2-furamidoxime, 5-nitrofurfurylidene diacetate and the urine of rats fed these compounds, were assayed for mutagenic activity in Salmonella typhimurium strains TA100 and TA100FR1. All the nitrofurans were mutagenic in the order: AF-2 and FANFT > nitrofurantoin > 5-nitro-2-furamidoxime > 5-nitrofurfurylidene diacetate > 5-nitro-2-furoic acid. Strain TA100 was more sensitive than TA100FR1 to the mutagenic influence of these nitrofurans. Only the urine of rats fed AF-2, FANFT and nitrofurantoin had mutagenic activity. Again, TA100 was more sensitive than TA100FR1. The mutagenicity of the urine was not increased by treatment with β-glucuronidase. AF-2, 2-amino-4-(5-nitro-2-furyl)thiazole (deformylated product of FANFT) and nitrofurantoin were excreted in the urine of rats fed these compounds; whereas the other nitrofurans were not excreted.     

A New Bipyrrole and Some Phenolic Constituents in Prunes (Prunus domestica L.) and Their Oxygen Radical Absorbance Capacity (ORAC)

Isolation and structural elucidation of prune constituents were performed and total 10 compounds were determined by NMR and MS analyses. A novel compound was identified to be 2-(5-hydroxymethyl-2′,5′-dioxo-2′,3′,4′,5′-tetrahydro-1′H-1,3′-bipyrrole)carbaldehyde, and 7 phenolic compounds were isolated from prunes for the first time. In addition, antioxidant activity of them was evaluated on the basis of the oxygen radical absorbance capacity (ORAC).     

Synthesis and in vitro evaluations of 6-(hetero)-aryl-imidazo[1,2-b]pyridazine-3-sulfonamide’s as an inhibitor of TNF-α production

Tumor necrosis factor-α is an important pro-inflammatory cytokine having a key role in hosts defensive process of immune systems and its over expression led to a diverse range of inflammatory diseases such as Rheumatoid arthritis, Cronh’s disease, psoriasis, etc. This paper describes our medicinal chemistry efforts on imidazo[1,2-b]pyridazine scaffold: design, synthesis and biological evaluation. By the introducing sulfonamide functionality at 3 positions and substituting 6 positions with (hetero)-aryl groups’, a small library of compounds was prepared. All synthesized compounds were screened for lipopolysaccharide (LPS) mediated TNF-α production inhibitory activity. Biological data revealed that the majority of the compounds of this series showed moderate to potent TNF-α production inhibitory activity. Compound 5u and 5v are the most potent compounds from the series with activity of IC₅₀?=?0.5?µM and 0.3?µM respectively. A short SAR demonstrates that 3-sulfonyl-4-arylpiperidine-4-carbonitrile moiety on imidazo[1,2-b]pyridazine showed better activity compared to the 3-(4-aryllpiperazin-1-yl) sulfonyl) in hPBMC assay. The molecular modeling studies revealed that the potent TNF-α production inhibitory activity 5v due to the extra stability of complex because of an extra pi-pi (π-π) stacking, hydrogen-bonding interactions.     

Novel pyrazole integrated 1,3,4-oxadiazoles: Synthesis,characterization and antimicrobial evaluation

A novel series of 2-(5-methyl-1,3-diphenyl-1H-pyrazol-4-yl)-5-phenyl-1,3,4-oxadiazoles 7(a–m) were synthesized either by cyclization of N′-benzoyl-5-methyl-1,3-diphenyl-1H-pyrazole-4-carbohydrazide 4a using POCl₃ at 120 °C or by oxidative cyclization of hydrazones derived from various arylaldehyde and (E)-N′-benzylidene-5-methyl-1,3-diphenyl-1H-pyrazole-4-carbohydrazide 5(a–d) using chloramine-T as oxidant. Newly synthesized compounds were characterized by analytical and spectral (IR, ¹H NMR, ¹³C NMR and LC–MS) methods. The synthesized compounds were evaluated for their antimicrobial activity and were compared with standard drugs. The compounds demonstrated potent to weak antimicrobial activity. Among the synthesized compounds, compound 7m emerged as an effective antimicrobial agent, while compounds 7d, 7f, 7i and 7l showed good to moderate activity. The minimum inhibitory concentration of the compounds was in the range of 20–50 μg mL⁻¹ against bacteria and 25–55 μg mL⁻¹ against fungi. The title compounds represent a novel class of potent antimicrobial agents.     

酸渣树中一个新的黄烷类化合物

从酸渣树(Carapaguianensis Aubl.)枝条的乙醇提取物中首次分离到7个化合物,通过波谱方法鉴定,它们分别是:()-epicatechin-3-O-(3",5"-di-O-methyl)gallate(1)、(-)-catechin(2)、sciadopitysin(3)、cl eomi scosin B(4)、photogedunin(5)、chi socheton compound F(6)和odoratone(7).其中,化合物1为新黄烷类化合物,2～7为首次从该植物中分离得到.补充了化合物5的13C-NMR谱数据.活性测定显示,化合物7对大菜粉蝶(Pieris brassicae)三龄幼虫具有较强的杀虫活性,化合物2有一定的活性,而正丁醇部分只有很弱的活性.     

Novel N-benzylpyridinium moiety linked to arylisoxazole derivatives as selective butyrylcholinesterase inhibitors: Synthesis,biological evaluation,and docking study

A novel series of N-benzylpyridinium moiety linked to arylisoxazole ring were designed, synthesized, and evaluated for their in vitro acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities. Synthesized compounds were classified into two series of 5a-i and 5j-q considering the position of positively charged nitrogen of pyridinium moiety (3- or 4- position, respectively) connected to isoxazole carboxamide group. Among the synthesized compounds, compound 5n from the second series of compounds possessing 2,4-dichloroaryl group connected to isoxazole ring was found to be the most potent AChE inhibitor (IC₅₀ = 5.96 µM) and compound 5j also from the same series of compounds containing phenyl group connected to isoxazole ring demonstrated the most promising inhibitory activity against BChE (IC₅₀ = 0.32 µM). Also, kinetic study demonstrated competitive inhibition mode for both AChE and BChE inhibitory activity. Docking study was also performed for those compounds and desired interactions with those active site amino acid residues were confirmed through hydrogen bonding as well as π-π and π-anion interactions. In addition, the most potent compounds were tested against BACE1 and their neuroprotectivity on Aβ-treated neurotoxicity in PC12 cells which depicted negligible activity. It should be noted that most of the synthesized compounds from both categories 5a-i and 5j-q showed a significant selectivity toward BChE. However, series 5j-q were more active toward AChE than series 5a-i.     

Potential Inhibitors of Angiogenesis. Part I: 3-(Imidazol-4(5)-ylmethylene)indolin-2-ones

The synthesis and pharmacological evaluation of new 3-(imidazol-4(5)-ylmethylene)indolin-2-ones, analogues of SU-5416, are reported. The final compounds 20-51 were obtained by Knoevenagel coupling between the substituted indolin-2-ones 1-15 and either the formylimidazole derivatives 16-18 or 2-formyl-3,5-dimethylpyrrole 19. Methylation at the nitrogen atom of the indolin-2-one and/or imidazole moities was carried out in the presence of the couple NaH/DMF. A Mannich reaction afforded the 1-dimethylaminomethyl derivatives 43 and 48. The antiangiogenic activity of these compounds was evaluated in a three dimensional in vitro rat aortic ring assay. In this test, compound 20 induced a decrease of angiogenesis comparable to that observed with SU-5416; the vascular density indexes at 1 μM were 30±18 and 22±4 % of control, respectively. The compounds were also evaluated, in an independent manner, as inhibitors of the human EGF-receptor tyrosine kinase activity. As expected, only minor activities were observed with four compounds, out of thirty-one, exerting inhibitory effects in the range of 40-55 % at 10 μM concentration.     

Synthesis,characterization and in vitro antimicrobial evaluation of new compounds incorporating oxindole nucleus

New compounds incorporating with the oxindole nucleus were synthesized via the reaction of substituted isatins [5-methyl-, 5-chloro- and 1-hydroxymethyl isatins] with different nucleophiles. The structures of the newly compounds were elucidated on the basis of FTIR, ¹H NMR, ¹³CMR spectral data, GC/MS and chemical analysis. Investigation of antimicrobial activity of the new compounds was evaluated using broth dilution technique in terms of minimal inhibitory concentration (MIC) count against four pathogenic bacteria and two pathogenic fungi. Most of the new compounds are significantly active against bacteria and fungi. MIC showed that compound (4a) possesses higher effect on Gram-positive bacteria Bacillus cereus than the selected antibacterial agent sulphamethoxazole, whereas compound (11c) possesses more activity against Gram-negative bacteria Shigella dysenterie.