海洋蓝藻 Okeania hirsute 中的次级代谢产物及其细胞毒活性

目的 研究从海洋蓝藻 Okeania hirsute 中分离鉴定次级代谢产物并进行细胞毒活性评价。方法 三个天然产物(1~3)的结构确定是通过广泛的波谱数据,包括一维和二维核磁及质谱等。结果 化合物3对人大细胞肺癌细胞NCI-H460显示出强的细胞毒活性,IC₅₀值为0.384 μg/mL。结论 这是首次从Okeania属蓝细菌中分离获得化合物1~3。这些天然产物的发现表明Okeania属蓝细菌可以作为发现新颖化合物的重要来源。     

两株大白鲨鳃部来源青霉生物碱及其生物活性研究r*

目的 探索大白鲨鳃部共附生真菌菌株 Penicillium sp. BP2T2 和 Penicillium sp. TBG2-2 的次生代谢产物。方法 利用柱色谱和高效液相色谱对发酵产物进行分离纯化,综合运用质谱、一维与二维核磁共振谱、比旋光测定和圆二色谱来鉴定化合物结构,采用结晶紫法评价化合物细胞毒活性,采用改良的微孔板法评价卤虫 Artemia salina 致死活性,采用纸片法评价其抗菌活性。结果 从菌株 Penicillium sp. BP2T2 发酵产物中分离鉴定到 1 个生物碱 Meleagrin （1）,从菌株 Penicillium sp. TBG2-2发酵产物中分离鉴定到3个生物碱,分别为fructigenine A （2）、（-）-cyclopenol （3） 和 viridicatol （4）。化合物 1、2 和 4 显示了对人结肠癌 HCT116 细胞株的细胞毒性,其半抑制浓度（IC50）分别为 5.1 μg/mL、40.5 μg/mL 和 59.0 μg/mL,并显示了对金黄色葡萄球菌或大肠埃希氏菌弱的抑菌活性,25 μg/片剂量下抑菌圈直径为7.5~8.0 mm。化合物 2 对卤虫的 24 h 半致死浓度为 125 μg/mL。结论 首次报道从鲨鱼鳃真菌中分离得到化合物 1~3 以及化合物 1、2 和 4 对 HCT116 细胞株的细胞毒性,鲨鱼鳃真菌作为研究较少的真菌资源在产生生物碱等次生代谢产物方面具有一定潜力。     

一株南海红树林底泥来源抗真菌放线菌No. 41-51发酵物的化学成分研究

目的 对一株来源于南海红树林底泥的抗真菌放线菌No. H 41-51发酵物中的化学成分进行研究。方法 对该菌株进行发酵培养,将发酵物进行离心分离成菌丝和菌液,菌液用乙酸乙酯萃取,并对具有活性的该乙酸乙酯部位进行成分分离和鉴定;发酵物菌丝体用95-80%乙醇提取,再用不同极性的溶剂萃取,对具有活性的石油醚部位进行成分分离和鉴定。整个提取和分离过程用纸碟片法进行活性追踪分离。通过硅胶柱、Sephadex LH-20柱色谱及HPLC制备方法分离纯化化合物样品,用NMR、MS等光谱方法,并结合文献数据比对鉴定化合物结构。 结果 分离得到14个化合物,并分别将其鉴定为邻苯二甲酸二丁酯(1)、二(2-乙基己基)苯-1,2-二甲酸酯(2)、3,3-二吲哚-2-羟基-丙醇(3)、环(苯丙-丙)二肽(4)、环(R-脯-S-苯丙)二肽(5)、环(S-脯-S-苯丙)二肽(6)、环(D-苯丙-L-异亮)二肽(7)、dankasterone(8)、4-hydroxy-17R-methylincisterol(9)、Calvasterol B(10)、Calvasterol A(11)、抗霉素 A_1a(12)、抗霉素 A_1b(13)和甘油醇-1-单油酸酯(14)。体外活性实验表明：化合物8~11对MCF-7、SF-268和NCI-H460细胞株表现出程度不等的细胞毒活性,化合物12和13表现出抗白色念珠菌活性。结论 菌株H 41-51发酵可产生多种不同结构类型和生物活性的次生代谢产物。化合物8~11对MCF-7、NCI-H460和SF-268细胞株具有细胞毒活性,化合物12、13具有抗白色念珠菌活性。     

溶剂及其体积分数对6种肺癌细胞的细胞毒作用

目的探索既能提高难溶性中药单体的溶解度,又对肿瘤细胞低毒或者无毒的有机溶剂及其体积分数窗口,阐明溶剂不同体积分数(volume fraction,VF)对模型化合物莪术醇药效作用的影响。方法采用四甲基偶氮唑蓝显色(MTT)法,研究二甲基亚砜(DMSO)和乙醇两种有机溶剂对人恶性肿瘤细胞A549、NCI-H460、NCI-H1299、NCI-1650、LTEP-a2和SPC-A1等细胞活力的影响和毒性作用,并进一步研究用不同体积分数乙醇配制的莪术醇对A549和NCIH460的细胞毒作用及其差异性。结果在48 h内,当DMSO体积分数(DMSO volume fraction,DVF)≤0.008时对A549无细胞毒作用;DVF≤0.004时对NCI-H1650无细胞毒作用;DVF≤0.002时对NCI-H460无细胞毒作用;然而,另外3种细胞NCI-H1299、LTEP-a2和SPC-A1细胞则在DVF<0.001时即表现出一定的细胞毒性。在48 h内,当乙醇体积分数(ethanol volume fraction,EVF)≤0.004时对A549无细胞毒作用;EVF≤0.002时对NCI-H460和NCI-H1650无细胞毒作用;当EVF≤0.001时对NCI-H1299无细胞毒作用;对LTEP-a2和SPC-A1细胞则在EVF<0.001时即表现有一定毒性作用。当EVF低于0.01时乙醇对A549和NCI-H460无细胞毒作用,以其配制的莪术醇溶液对细胞活力的影响仅代表莪术醇的药效作用,由于EVF 0.01的乙醇比EVF0.001的乙醇具有更好的助溶作用,又具有细胞毒作用,两细胞株细胞活力差异极大,分别从93.84%和95.67%降至35.35%和36.41%。结论 A549、NCI-H1650可以首选DMSO作为溶剂,NCI-H1299可以首选乙醇作为溶剂,NCI-H460可选择其中任一溶剂,而LTEP-a2和SPC-A1则不宜以DMSO或者乙醇作为溶剂。     

中国南海刺柳珊瑚 Echinogorgia rebekka 中倍半萜氢醌糖苷类化合物及其生物活性△

目的 对中国南海刺柳珊瑚 Echinogorgia rebekka 中倍半萜氢醌糖苷类化合物进行分离鉴定,并测定其生物活性。方法 运用正反相硅胶柱层析和半制备高效液相色谱等分离手段对化合物进行分离和纯化;运用 NMR 和 MS 等现代波谱方法,并结合相关文献,对化合物的结构进行鉴定;运用细胞毒活性和抗菌活性模型对化合物的生物活性进行评价。结果 从该种柳珊瑚中分离获得了 4 个倍半萜氢醌糖苷类化合物 euplexides A–D (1–4)。化合物 1–4 对肿瘤细胞 HeLa, P388和 A549均显示出中等强度的抑制活性,化合物 2 对恶臭假单胞菌 Pseudomonas putida (MIC = 0.78 μM) 和巴西诺卡菌 Nocardia brasiliensis (MIC = 0.78 μM) 的抑制活性与阳性药环丙沙星相当。结论 化合物 1–4 均为首次从 Echinogorgia 属柳珊瑚中分离得到,具有作为抗菌药的研究价值。     

盐碱地真菌棘孢曲霉Aspergillus aculeatus的化学成分研究△﹡

目的 对一株具有抗肿瘤活性的滨州盐碱地真菌棘孢曲霉Aspergillus aculeatus的活性次级代谢产物进行研究。方法 采用硅胶柱层析、LH-20凝胶色谱和半制备高效液相色谱分离纯化代谢产物,通过理化性质、波谱手段结合X-ray单晶衍射技术鉴定化合物,采用MTT法测试所得化合物对肿瘤细胞A549的增殖抑制活性。结果 从棘孢曲霉Aspergillus aculeatus中分离得到8个化合物(1～8),分别鉴定为5-Hydroxymethyl-2-furoic acid (1)、Isorhodoptilometrin (2)、Secalonic acid F (3)、Emodin (4)、Protocatechuic acid (5)、Altechromones B (6)、Cyclo (L-Tyr-L-Leu) (7)和Cyclo (L-Phe-L-Ala) (8)。化合物3对A549细胞具有一定的增殖抑制活性,其IC50值为14.9 μM。结论 首次研究了来源于盐碱地真菌棘孢曲霉Aspergillus aculeatus的活性次级代谢产物,并首次报道secalonic acid F对A549肿瘤细胞增殖抑制活性。     

海绵共附生真菌Penicillium janthinellum的次生代谢产物研究

目的 研究海绵共附生真菌Penicillium janthinellum LZDX-32-1中的活性次生代谢产物。方法 利用硅胶、半制备HPLC等色谱学方法对菌株发酵提取物进行分离纯化，利用UV、NMR、MS等波谱学分析及文献数据比对确定了化合物的结构，采用MTT法检测了化合物的人肿瘤细胞毒性。结果 分离鉴定了5个化合物，分别为okaramines H（1）、okaramines J（2）、fumitremorgin B（3）、verruculogen（4）、paspaline（5）。细胞毒活性测试发现在浓度为10μM条件下，化合物5对A549、HCT-8和MCF-7三种细胞具有一定细胞毒活性（抑制率分别为70.45%、67.03%和88.54%）。结论 从海绵共附生真菌P. janthinellum LZDX-32-1中分离鉴定了5个生物碱类化合物，其中化合物1-4为首次从该种真菌中分离得到，化合物5具有肿瘤细胞毒性。     

一株六放珊瑚来源真菌Exserohilum sp.中活性天然产物研究

目的 研究六放珊瑚来源的真菌Exserohilum sp.中活性天然产物。方法 综合运用硅胶柱层析、Sephadex LH-20凝胶柱层析和半制备型HPLC等分析和分离技术分离纯化化合物,利用核磁、质谱等现代波谱分析方法对化合物进行结构鉴定,以致病真菌、细菌和海洋污损细菌为活性模型对化合物进行活性评价。结果从Exserohilum sp.中分离鉴定了4个天然产物 (1–4),化合物3对致病真菌Candida albicans和海洋污损细菌Pseudomonas aeruginosa显示中等抑制活性,MIC 分别为16.33 和32.65 μg/mL。结论 从海洋来源的真菌中分离获得具有生物活性的天然产物1–4,化合物3 对致病真菌和海洋污损细菌显示中等抑制活性。     

十字孢碱产生菌筛选及其肿瘤细胞毒活性产物研究

目的 寻找海洋放线菌来源的十字孢碱产生菌,筛选获得目标菌株,分离鉴定其具有吲哚咔唑母核的产物。方法 以十字孢碱在λ_max290 nm附近尖锐的特征紫外吸收为标准,利用HPLC-UV对不同来源的海洋放线菌的发酵提取物进行定向筛选,筛选出十字孢碱产生菌,利用16S rRNA序列对筛选得到的菌株进行种属鉴定。通过比较各产生菌的十字孢碱类产物的丰度,选择能够产生更多该类化合物的菌株进行规模化发酵,并采用硅胶柱层析、Toyopearl HW-40F凝胶柱层析、高效液相色谱(HPLC)等方法对其发酵提取物进行分离、纯化,运用质谱(MS)、核磁共振(NMR)等技术鉴定化合物的结构。采用CCK-8法测定化合物的细胞毒活性。 结果 从42株放线菌中筛选出12株十字孢碱的产生菌,其中Streptomyces sp. OUCMDZ-5380的产物最为丰富,从该菌株发酵提取物中分离鉴定了3个吲哚咔唑类化合物,分别为K252c (1)、十字孢碱 (2)和4"-O-去甲基十字孢碱 (3)。细胞毒活性测试表明,化合物3对12株肿瘤细胞株的IC₅₀值达到纳摩尔、介于0.0003–0.623 μM之间。 结论 筛选得到12株主产十字孢碱的海洋放线菌,从OUCMDZ-5380的发酵产物中鉴定了3个吲哚咔唑类化合物,化合物3具有广谱的肿瘤细胞毒活性(半数抑制浓度IC₅₀为0.0003–0.623 μM.)和对内部串联复制突变的人急性髓细胞性白血病细胞MV4-11的选择性(相对于人正常细胞株L-02和其它人肿瘤细胞株的选择指数分别为1120和50–2080)。     

南海沉积环境来源真菌Eurotium sp.SCSIO F452的次生代谢产物研究

目的对1株南海沉积环境来源真菌的次生代谢产物进行分离、鉴定及活性研究。方法采用溶剂萃取、硅胶柱层析、凝胶柱层析等方法对真菌Eurotiumsp.SCSIO F452的发酵产物进行化学分离,通过NMR、MS等波谱学技术并参阅文献进行化合物结构鉴定,采用SRB法评价化合物的细胞毒活性。结果从菌株SCSIO F452中分离鉴定6个单体化合物,分别为:isodihydroauroglaucin(1)、flavoglaucin(2)、tetrahydroauro-glaucin(3)、2-(1,1-dimethyl-2-propen-1-yl)-1H-indole-3-carboxaldehyde(4)、neoechinulin A(5)和methyl lino-leate(6)。化合物1-5对4种肿瘤细胞系表现出不同强度的细胞毒活性。结论苯甲醛衍生物1-3是真菌SC-SIO F452的优势代谢产物,细胞毒活性较强,具有潜在的研究价值。     

Hemorrhagic activity of the Duvernoy''s gland secretion of the xenodontine colubrid Philodryas patagoniensis from the north-east region of Argentina

Colubrid snakes belonging to Philodryas genus, widespread all over South America, bring about lesions (swelling, ecchymosis, transient bleeding from the bite site punctures), that are similar to those produced by Bothrops species (yarará). In the present work we began the characterization of Philodryas patagoniensis venom. We examined if this venom produces hemorrhagic lesions as those observed in victims bitten by Philodryas olfersii. Hemorrhagic, proteolytic and fibrinogenolytic activities were evaluated, and histological observations in samples of gastrocnemius muscle were carried out. Inhibition studies were carried out in metal chelator (ethylenediaminetetraacetic acid) presence. Our results show a small Minimum Hemorrhagic Dose (MHD=0.035 μg) and a high proteolytic activity (143 U/mg), and prove the capacity of this venom to degrade fibrinogen in vitro rendering it unclottable by thrombin, supporting the presence of proteases, principally metalloproteases, in P. patagoniensis venom that are able to alterate the vascular wall and degrade fibrinogen, being both activities responsible of a high hemorrhagic activity.     

Evaluation of the Antimicrobial Activity of Ammoniacum Gum from Dorema ammoniacum

Antimicrobial activity of the dichloromethane-methanol (1 : 1) extract of ammoniacum gum (from Dorema ammoniacum D. Don) was evaluated against 14 microorganisms which included seven Gram-positive bacteria (Bacillus cereus, Bacillus pumilus, Bacillus subtilis, Micrococcus luteus, Staphylococcus epidermidis, Staphylococcus aureus and Streptococcus faecalis), four Gram-negative bacteria (Escherichia coli, Pseudomonas aereuginosa, Klebsiella pneumoniae and Bordetella bronchiseptica), one yeast (Saccharomyces cereviseae) and two fungi (Aspergillus niger and Candida albicans). The extract of ammoniacum gum exhibited a of broad spectrum antimicrobial activity by inhibiting all the seven Gram-positive bacterium, one Gramnegative bacterium, one yeast and one fungus, with a minimum inhibitory concentration (MIC) of 40µg/ml. To overcome the solubility problem often faced when herbal extracts are added to aqueous medium, we employed a modified broth method where the broth cultures were agitated at 150 rpm in an orbital shaking incubator. This method reduced the MIC of the extract considerably, to 5-20µg/ml, against B. bronchiseptica, S. aureus and S. epidermidis.     

Investigation of the antimicrobial and anticancer activity of aminonaphthoquinones

In this study, we report on the inhibitory activity of synthesized aminonaphthoquinones against two bacterial and one fungal species to determine their antimicrobial properties. A minimum inhibitory concentration (MIC) of 7.8 μg/mL was obtained against the fungus, Candida albicans, which was better than that of Amphotericin B (MIC = 31.25 μg/mL). Escherichia coli (Gram -), was inhibited at a MIC of 23.4 μg/mL and Staphylococcus aureus (Gram +) at a MIC of 31.3 μg/mL. The aminonaphthoquinones were also screened against HCT116 colon, PC3 prostate and HepG2 liver cancer cell lines to evaluate their cytostatic effects. They had potent activity (GI₅₀ = 5.87–9.90 μM) which was about three-6-fold better than that of parthenolide (GI₅₀ = 25.97 μM) against the prostate cancer cell line. These compounds were generally more selective for cancer cells than for normal human lung fetal fibroblasts (WI-38).     

SV预防血栓药理学作用及其机制研究探讨

血栓栓塞性疾病是人类首要致死病因,临床预防血栓药物均存在出血危险。选择性的内源性凝血途径抑制剂已成为低出血倾向预防血栓药物研发的重点方向。SV是以天然岩藻糖化糖胺聚糖(FG)为先导化合物经结构优化获得的选择性内源性凝血因子Xase抑制剂(IC50=30 nm),其抑制内源性凝血活性与依诺肝素相当,在等效抗凝血剂量的倍数剂量下出血倾向显著降低。本文探讨低出血倾向的SV预防血栓药理学作用及其机制,为选择性Xase抑制剂及其新药发现研究提供参考依据。     

Antipyretic, analgesic and anti-inflammatory effects of delta 9-tetrahydrocannabinol in the rat

The effects on body temperature produced by graded doses of Δ⁹-tetrahydrocannabinol (Δ⁹-THC) and phenylbutazone were compared in both normal and pyretic rats. Dose related hypothermic responses were produced by the oral administration of Δ⁹-THC in normal animals. Moreover, Δ⁹-THC significantly reduced elevated temperatures in yeast-induced pyretic rats to near normal levels at doses which exhibited little hypothermic activity in normal rats. The oral antipyretic potency of Δ⁹-THC was approximately 2 times that of phenylbutazone. The comparative oral antinociceptive activity of Δ⁹-THC and selected narcotic and non-narcotic analgesics was determined by the increase in response latency to pressure applied to normal and yeast-inflamed paws. Δ⁹-THC administered orally was essentially inactive at dose levels below those producing pronounced central nervous system depression. The oral anti-inflammatory efficacy of Δ⁹-THC was compared to phenylbutazone and acetylsalicylic acid. Δ⁹-THC was ineffective in inhibiting carrageenin-induced edema of the rat paw following acute or chronic administration.     

Chemical constituents and bioactivities of the liposoluble fraction from different medicinal parts of Crocus sativus

Context: Crocus sativus Linn. (Iridaceae), commonly known as saffron, becomes more and more popular due to its versatile biological and medicinal properties. At present, studies mainly focus on the traditional medicinal part, the saffron stigma, with less attention to the other parts of saffron, such as the perianth, the stamen, and the corm, which are high yield compared to the stigma and also possess various pharmacological effects.

Objective: To determine the chemical compositions, antifungal, cytotoxic, and antioxidant activities of the ether fractions from the stamen, perianth, and stigma of saffron.

Materials and methods: The chemical constituents of the ether fractions from different parts of saffron were investigated by gas chromatography/mass spectrometry. Several pathogenic fungi isolates and tumor cell lines were employed to evaluate the antifungal and cytotoxic activities of these three ether fractions. 1,1-Diphenyl-2-picrylhydrazyl assay was used to determine the free radical-scavenging activity.

Results: The ether fractions composition of the three C. sativus parts are different from each other, but lauric acid, hexadecanoic acid, 4-hydroxydihydro-2(3H)-furanone, and stigmasterol were the common constituents shared by all the three fractions. The stamen ether fraction displayed the strongest antifungal and cytotoxic activities, whereas both of the saffron stamen and perianth ether fractions exhibited significant antioxidant activities.

Discussion and conclusion: These findings demonstrate that the saffron stamen and perianth possess significant antifungal, cytotoxic, and antioxidant activities as well as the stigma, though not to the same extent, prompting us to expand the medicinal resource and make best use of this valuable plant.     

Antioxidant and vasorelaxant activities of the Turraeanthus africanus methanol extract

The present studies evaluate the antioxidant and vasorelaxant activities of the methanol stem bark extract of Turraeanthus africanus (Welw.) Pellegr. (Meliaceae). The antioxidant property investigated through the DPPH scavenging activity showed that the methanol extract from the stem bark of T. africanus was active with an inhibition rate of 72.47% at 500 μg/mL and IC₅₀ of 29.2 μg/mL. This extract (0.1–700 μg/mL) induced a concentration-dependent relaxation of guinea-pig aortic rings precontracted with noradrenaline or KCl, with a maximum relaxation reaching 87.55 and 97.42%, respectively, at a concentration of 700 μg/mL. The extract showed no significant effect on the electrically induced contraction of the guinea-pig papillary muscle. The results of this study indicate that the extract from the stem bark has interesting antioxidant and vasorelaxant properties and represents a potential source of medicine for the treatment of cardiovascular diseases.     

Evaluation of the antimalarial and antileishmanial activity of plants from the Greek island of Crete

Different parts of 65 plant species from the Greek island of Crete have been extracted and the 249 extracts obtained have been investigated for in-vitro antiprotozoal activity. Their activity against chloroquine-sensitive (D6) and resistant (W2) strains of Plasmodium falciparum and Leishmania donovani promastigotes was determined. Their cytotoxicity on a mammalian kidney fibroblast (Vero) cell line was also tested. Dichloromethane and methanol extracts of Berberis cretica and methanol extracts of Cytinus hypocistis subsp. hypocistis, C. hypocistis subsp. orientalis, and C. ruber had significant activity against both strains of P. falciparum (IC₅₀<10 μg mL⁻¹). Dichloromethane extracts of Eryngium ternatum, Origanum dictamnus, and Origanum microphyllum, and the methanolic extract of Eryngium amorginum had significant activity against Leishmania donovani (IC₅₀<10 μg mL⁻¹). None of the extracts was cytotoxic.     

Antimicrobial and antineoplastic activities of agelasine analogs modified in the purine 2-position

Agelasines are 7,9-dialkylpurinium salts found in marine sponges (Agelas sp.), which display a variety of antimicrobial and cytotoxic effects. We have synthesized simplified agelasine analogs modified in the purine 2-position and examined their antimicrobial and anticancer activities. The compounds were screened against Staphylococcus aureus, Escherichia coli, Mycobacterium tuberculosis, Candida krusei, and Candida albicans, protozoa causing tropical diseases (Plasmodium falciparum, Leishmania infantum, Trypanosoma cruzi, and Trypanosoma brucei), a panel of human cancer cell lines (U-937 GTB, RPMI 8226/s, CEM/s, and ACHN) as well as VERO and/or MRC-5 cells. The results indicate that the introduction of a methyl group in the purine 2-position is beneficial for antimycobacterial and antiprotozoal activity, and that amino groups may enhance activity against several cancer cell lines.