续随子中千金二萜烷化合物抑制人妇科肿瘤细胞增殖活性的研究

目的检测大戟科植物续随子中分离纯化的3种千金二萜烷化合物Euphorbia Factor L1(1)、Euphorbia Factor L2(2)和Euphorbia Factor L3(3)对子宫颈癌、子宫内膜癌、卵巢透明癌和卵巢囊腺癌等5种人妇科肿瘤细胞增殖的影响,探讨了千金二萜烷化合物结构与抑制肿瘤细胞增殖活性之间的构-效关系。方法利用四唑盐(MTT)比色法测定阳性对照顺铂和3种化合物对子宫颈癌、子宫内膜癌、卵巢透明癌和卵巢囊腺癌等5种人妇科肿瘤细胞增殖的影响。结果发现Euphorbia Factor L1(1)和Euphorbia Factor L3(3)对子宫颈癌细胞的增殖具有抑制作用,EC50分别为16.74和1.01μmol.L-1;Euphorbia Factor L3(3)对人卵巢透明癌和人卵巢囊腺癌细胞的增殖显示中等程度的抑制活性;Eu-phorbia Factor L2(2)即使在100μmol.L-1的高浓度下,对5种肿瘤细胞的增殖不显示抑制活性。结论 Euphorbia Fac-tor L3(3)对人妇科肿瘤细胞的增殖显示抑制活性,推测千金二萜烷化合物抑制肿瘤细胞增殖活性与母核上的环外双键及邻位取代基有关。     

倍半萜内酯及有关化合物的抗高血脂活性

作者研究比较了18种倍半萜内酯及有关化合物的抗小鼠高血脂活性。结果发现其中堆心菊内酯(Helenalin),细叶土木香苦素(Tenulin),2.3-环氧异细叶土木香苦素,异细叶土木香苦素,2.3-环氧异细叶土木香苦素,     

异土木香内酯体外抗病毒作用与机制研究

目的 研究异土木香内酯的体外抗病毒作用及机制。方法 CCK-8法检测0.125、0.500、1.000、2.000、4.000、8.000、16.000、32.000、64.000 μmol·L^-1的异土木香内酯处理24 h对人非小细胞肺癌细胞系（A549）细胞活力的影响;表达绿色荧光蛋白的水疱性口炎病毒（VSV-eGFP）以0.02的感染复数（MOI）感染A549细胞制备模型,流式细胞术检测共同孵育12 h的异土木香内酯5、10、20 μmol·L^-1对GFP阳性细胞率的影响;VSV感染（MOI=0.1） A549细胞,Western blotting检测异土木香内酯处理16 h对VSV病毒G蛋白表达的影响;分别使用甲型流感病毒（H1N1,MOI=0.1）、脑心肌炎病毒（EMCV,MOI=0.1）感染A549细胞,同时给药共同孵育8 h后,实时荧光定量PCR （qRT-PCR）检测土木香内酯对病毒RNA表达的影响;异土木香内酯分别以预处理12 h、病毒吸附过程中给药2 h、病毒吸附后给药10 h 3种不同方式给药,通过流式细胞术检测其对VSV-eGFP在A549细胞中复制的影响;异土木香内酯20 μmol·L^-1处理胚胎成纤维（MEF）细胞12 h,进行转录组测序分析;异土木香内酯5、10、20 μmol·L^-1处理MEF细胞12 h,qRT-PCR检测干扰素α1（Ifna1）、干扰素β1（Ifnb1）、干扰素诱导蛋白44（Ifi44）、干扰素刺激基因15（Isg15）的mRNA表达。结果 异土木香内酯对A549细胞的半数抑制浓度（IC₅₀）为51.01 μmol·L^-1;与模型组比较,流式结果显示异土木香内酯显著减少VSV-eGFP阳性细胞率（P<0.001）,但对病毒的吸附过程没有影响,药物预处理及吸附后给药显著抑制VSV病毒复制（P<0.01、0.001）;qRT-PCR结果显示异土木香内酯显著降低H1N1、EMCV病毒的mRNA水平（P<0.001）;Western blotting结果显示异土木香内酯显著抑制VSV的G蛋白表达（P<0.001）;转录组测序分析和qRT-PCR结果表明,异土木香内酯促进I型干扰素通路的激活,并诱导Ifna1、Ifnb1、Ifi44、Isg15（P<0.001）表达。结论 异土木香内酯通过促进基于干扰素通路的抗病毒天然免疫激活,发挥抑制病毒复制的作用。     

果旭阿汤中土木香内酯和异土木香内酯的含量测定

［摘要］目的建立同时测定藏药果旭阿汤中土木香内酯和异土木香内酯含量的方法。方法通过优化提取方法,用气相色谱（GC）法同时测定藏药果旭阿汤中藏木香有效成分土木香内酯和异土木香内酯的含量。结果土木香内酯和异土木香内酯均在0.1～1.0 mg&#8226;mL-1 (n=5）范围内呈良好线性关系。土木香内酯回收率为97.25% (n=6),RSD=0.93%;异土木香内酯回收率为97.42% (n=6), RSD=0.90%。结论该方法可用于藏药果旭阿汤中土木香内酯和异土木香内酯的含量同时快速、准确测定。     

木果楝内酯化合物抑制人肺肿瘤细胞增殖活性及作用机制的研究

目的检测3种木果楝内酯化合物(Xylocarpin H,Xy-logranatin C和Xylomexicanin A)对人肺肿瘤细胞增殖的抑制作用,并进一步探讨其可能的作用机制。方法 MTT比色法检测细胞增殖抑制率;采用Western blot分析与凋亡相关的蛋白表达变化。结果 Xylogranatin C对人非小细胞肺癌细胞(A549、RERF-LC-jk和QG-56)以及人小细胞肺癌细胞(PC-6和QG-90)5种实验用细胞的增殖均显示较强的抑制活性;Western blot结果显示由Xylogranatin C处理的A549细胞中的p53、Bax和caspase-3的表达与对照组相比有明显的增加(P<0.05)。结论 Xylogranatin C具有较强的抑制人肺肿瘤细胞增殖作用,其作用机制可能与其诱导肿瘤细胞凋亡有关。     

重组人内抑素的抗肿瘤活性

目的观察重组人内抑素抗肿瘤活性及对血管内皮细胞增殖的抑制作用。方法用人癌裸鼠移植性肿瘤及小鼠移植性肿瘤模型对重组人内抑素进行抗肿瘤药效学观察,用MTT法观察其对血管内皮细胞及肿瘤细胞增殖的抑制作用。结果重组人内抑素,可明显抑制人胃癌BGC803和人乳腺癌B37裸鼠移植性肿瘤的生长,对小鼠肝癌H22实体瘤亦呈一定的抑制作用。MTT试验结果显示重组人内抑素可抑制人胎脐静脉血管内皮细胞ECV304的增殖,对人结肠癌HCT-8等肿瘤细胞的增殖无影响。结论重组人内抑素具有较强的抗肿瘤作用,其作用机理可能与抑制血管内皮细胞增殖、抑制肿瘤新生血管形成有关。     

HPLC法同时测定利胆解毒胶囊中异土木香内酯和土木香内酯的含量

目的：建立高效液相色谱法（HPLC）同时测定利胆解毒胶囊中异土木香内酯和土木香内酯含量的方法。方法：采用Diamonsil Plus C18色谱柱（4.6 mm×250 mm，5 μm），流动相为乙腈-0.05%磷酸溶液（50:50），流速1.0 mL·min-1，检测波长210 nm，柱温30.0 ℃。结果：异土木香内酯、土木香内酯线性范围分别为0.044～1.325 μg（r=0.999 98）和0.038～1.149 μg（r=0.999 97）；平均回收率分别为98.55%和98.68%，RSD分别为0.86%和0.89%。测定4批样品，结果每粒含异土木香内酯和土木香内酯的含量分别在0.64～1.08 mg和0.43～0.68 mg之间。结论：该方法操作简便、重复性好、结果准确，可作为利胆解毒胶囊质量的控制方法。     

含嘌呤骨架的EGFR双突变体抑制剂的合成及其活性评价

目的设计、合成含嘌呤骨架的EGFR双突变体抑制剂,并评价其抗肺癌细胞增殖活性。方法以2,4-二氯-5-硝基嘧啶和相应的胺为起始原料,合成中间体1a～1d;以2-甲氧基-4-氟-5-硝基苯胺为原料,经氨基保护、亲核取代、还原、亲核取代、脱保护基后,与中间体1a～1d进行亲核取代反应,再经还原、亲核加成-消除反应合成目标化合物TM1～TM4。采用MTT法评价目标化合物对3种人非小细胞肺癌细胞(A549、HCC827、H1975)增殖抑制活性;采用ADP-Glo法评价目标化合物抑制EGFR~(L858R/T790M)激酶活性。结果与结论合成了4个未见文献报道的新化合物,其结构均经1 H-NMR和LC-MS谱确证。体外活性测试结果表明,TM1、TM2、TM4显示出较强的抗H1975和HCC827细胞增殖活性;4个目标化合物对EGFR突变细胞(HCC827和H1975)的抑制活性均强于对EGFR野生型(WT)的细胞(A549);TM1和TM2抑制EGFR~(L858R/T790M)的IC50值分别为2.5、 2.0nmol·L~(-1),与阳性药AZD9291(3.5nmol·L~(-1))相当。以嘌呤为骨架,设计合成的目标化合物对EGFR双突变的细胞有较强的抑制活性;目标物TM1和TM2对EGFR~(L858R/T790M)激酶显示出强的抑制作用。     

凋亡抑制蛋白抑制剂的设计合成与抗肿瘤活性评价

目的设计并合成具有抗肿瘤活性的小分子凋亡抑制蛋白广泛性抑制剂。方法运用基于靶点结构的药物设计策略改造前期得到的苗头化合物,设计合成新结构化合物,并进行多靶点的凋亡抑制蛋白家族（IAPs）结合实验和多个细胞系的肿瘤细胞增殖抑制活性评价。结果合成了25个新结构化合物,部分表现出较强的IAPs抑制活性和多种肿瘤细胞增殖抑制能力（微摩尔级别）,Ⅱc系列化合物显示出较强的ciAP1选择性（〉1000）。结论五元杂环烷当中的4s-甲基是化合物对两类IAPs产生选择性差异的结构基础。     

5-氟尿嘧啶衍生物的合成与抗肿瘤活性研究

目的寻找新型、高效的5-氟尿嘧啶衍生物类抗肿瘤化合物。方法设计、合成4个5-氟尿嘧啶衍生物,利用MTT方法测定这些化合物对4株肿瘤细胞的增殖抑制活性。结果 4个5-氟尿嘧啶衍生物均显示了较好的肿瘤细胞增殖抑制活性。结论 4个目标化合物对肿瘤细胞有较好的抑制活性,值得进一步研究。     

Cytotoxic triterpene and sesquiterpene lactones from Elephantopus mollis and induction of apoptosis in neuroblastoma cells

Activity-guided fractionation of root and leaf extracts from Elephantopus mollis led to the isolation of a new triterpene (1) and a new sesquiterpene lactone (2) together with five known sesquiterpene lactones (3-7). The structures of compounds 1 and 2 were determined based on their spectroscopic data. The cytotoxic activity of the isolated compounds was evaluated against neuroblastoma B104 cells. The sesquiterpene lactone-type compounds 2-7 were highly cytotoxic. Among these, compound (5) was the most cytotoxic and induced apoptosis of neuroblastoma B104 cells in a dose- and time-dependent manner. No significant difference was observed for cytotoxicity of compound 5 towards all 3 cell lines tested.     

Chemical constituents from the roots of Tripterygium wilfordii and their cytotoxic activity

In our ongoing search for bioactive constituents, a new sesquiterpene polyol ester, named triptersinine U (1), together with five known triterpenes (2–6) and seven sesquiterpene pyridine alkaloids (7–13), were isolated from the roots of Tripterygium wilfordii Hook. f. Their chemical structures were elucidated using extensive spectroscopic analyses, including 1D and 2D NMR, and HRESIMS, as well as comparison with previously reported data. Cytotoxic activities of all compounds 1–13 were evaluated against six human tumor cell lines (HepG2, Hep3B, Bcap37, U251, MCF-7 and A549) using the MTT in vitro assay. The results showed that triterpenes exhibited moderate cytotoxic activities toward the tested cell lines.     

A new sesquiterpene lactone from Tsoongiodendron odorum Chun.

A new sesquiterpene lactone (1) was obtained from the cytotoxic fraction of 95% ethanol extract of root barks of Tsoongiodendron odorum Chun together with two known sesquiterpene lactones, costunolide (2) and parthenolide (3). The structure of 1 was elucidated as 5alpha, 6alpha, 7beta, 10beta- 11alpha, 13-dihydro-4(15)-eudesmene-12, 6-olide on the basis of chemical and spectral evidence including X-ray diffraction analysis. Costunolide showed cytotoxic activity against human leukemia (HL-60) cell line. Parthenolide showed promising cytotoxic activities in vitro against HCT-8, Bel-7402, SKOV3, KB, HELA and EJ cell lines. Also, the cytotoxic ethyl acetate fraction of ethanol extract of the root barks from which three chemical components were isolated showed promising cytotoxic activities in vitro against KB, BGC-823, Bel-7402, HCT-8, HL-60 cell lines.     

Isoalantolactone, a sesquiterpene lactone, induces apoptosis in SGC-7901 cells via mitochondrial and phosphatidylinositol 3-kinase/Akt signaling pathways

Isoalantolactone, a sesquiterpene lactone, possesses anti-fungal as well as cytotoxic properties. In this study, the effects of Isoalantolactone on cell viability, cell cycle, and apoptosis were investigated in human gastric adenocarcinoma SGC-7901 cells. The results demonstrated that Isoalantolactone induced morphological changes and decreased cell viability. Subsequently, we found that Isoalantolactone induced G2/M and S phase arrest, which was associated with a decrease in the expression level of cyclin B1. Apoptosis triggered by Isoalantolactone was visualized using propidium iodide (PI) and Annexin V-FITC/PI staining. Isoalantolactone-induced apoptosis of SGC-7901 cells was associated with the dissipation of mitochondrial membrane potential (ΔΨ _m) that was due to the down-regulation of Bcl-2 and up-regulation of Bax that led to the cleavage of caspase-3. Additionally, it was found that Isoalantolactone was involved in the inhibition of phosphorylation of PI3K/Akt. Isoalantolactone-induced cytotoxicity and apoptosis of SGC-7901 cells involve mitochondria-caspase and PI3K/Akt dependent pathways, which gives the rationale for in vivo studies on the utilization of Isoalantolactone as a potential cancer therapeutic compound.     

Cytotoxic furanogermacranolides from the flowers of Helianthus angustifolius

The dichloromethane extract of the flowers of Helianthus angustifolius L. (Asteraceae, Heliantheae) was investigated in vitro for its cytotoxic activity using human cancer cell lines: CCRF-CEM leukemia, MDA-MB-231 breast cancer, U251 glioblastoma, HCT 116 colon cancer cells, and the human lung fibroblast cell line MRC-5. Cytotoxicity-guided fractionation led to the isolation of four related heliangolide-type sesquiterpene lactones. The structures were elucidated by means of NMR spectroscopy and high-resolution mass spectrometry. Of the investigated compounds, 8-methacrylyl-4,15-iso-atriplicolide (1) showed the highest activity against all tested cancer cell lines with IC?? values ranging from 0.26?±?0.01?μM for CCRF-CEM cells to 4.22?±?0.26?μM for MRC-5 cells.     

Synthesis and Evaluation of Biological and Antitumor Activities of Tetrahydrobenzothieno[2,3‐d]Pyrimidine Derivatives as Novel Inhibitors of FGFR1

A series of tetrahydrobenzothieno[2,3‐d]pyrimidine derivatives were designed, synthesized, and evaluated as inhibitors of FGFR1. These analogs were synthesized via Gewald's reaction under mild conditions. The structures of the synthesized compounds were characterized by spectroscopic data (IR, ¹H NMR and MS). Their antitumor activities were evaluated against H460, A549 and U251 cell lines in vitro. Results revealed that the tested compounds showed moderate antitumor activities. Structure–activity relationship analyses indicated that compounds with an aromatic ring substituted in the C‐2 position or with larger molecules such as 3g , 4c , and 7 were more effective than others. The compound, 3g (78.8% FGFR1 inhibition at 10 μ m ), was identified to have the most potent antitumor activities, with IC₅₀ values of 7.7, 18.9, and 13.3 μ m against the H460, A549, and U251 cell lines, respectively. Together, the results suggested that tetrahydrobenzothieno[2,3‐d]pyrimidine derivatives may serve as a potential agent for the treatment of FGFR1‐mediated cancers.     

Effects of sesquiterpene lactones on mitochondrial oxidative phosphorylation

1. The toxic plant sesquiterpene lactones, helenalin, hymenoxon, mexicanin-E, tenulin, dihydrogriesenin, and psilotropin which were isolated from Helenium, Hymenoxys, and Geigeria spp markedly inhibited "state 3" respiration in mouse hepatic mitochondria. With the exception of dihydrogriesenin and psilotropin, all the other sesquiterpene lactones stimulated "state 4" respiration. 2. The sesquiterpene lactones also stimulated ATPase activity in the presence of Mg2+ ions and caused mitochondrial swelling in buffer solutions containing magnesium, sodium, ammonium and potassium chloride salts. 3. The number of alkylating sites present in a sesquiterpene lactone appears to be related to the inhibitory activity on mitochondrial oxidative phosphorylation, however, the structure-activity relationship of these lactones is not clear at present. 4. Mitochondria prepared from the liver of ethoxyquin hydrochloride-fed mice were less susceptible to sesquiterpene lactone-mediated inhibition of mitochondrial oxidative phosphorylation.     

Two new eudesmanolides from Inula racemosa

Two new eudesmane-type sesquiterpene lactones were isolated from the roots of Inula racemosa and their structures were elucidated as 3β-hydroxy-11α,13-dihydroalantolactone (1) and 11α-hydroxy-eudesm-5-en-8β,12-olide (2). Their cytotoxic activities against five human cancer cell lines had been tested and compound 2 exhibited weak cytotoxic activity against BEL-7402 and HCT-8 cell lines. The anti-inflammatory activities were also tested, but neither of them showed any activities.     

Perturbation of glutathione status and generation of oxidative stress in mouse skin following application of contact allergenic sesquiterpene lactones and isothiocyanates

1. The sensitizing or non-sensitizing status of selected sesquiterpene lactones and isothiocyanates was confirmed in mouse by open epicutaneous application.

2. Glutathione status of mouse skin was determined 12 h after lactone/isothiocyanate application; glutathione S-transferase activity also was determined 12 h after lactone application.

3. NAD(P)H utilization by rat liver microsomal preparations exposed to the sesquiterpene lactones and isothiocyanates was measured.

4. A correlation was observed between sensitizing status and the ability to perturb glutathione status, to induce glutathione S-transferase activity, and to stimulate NAD(P)H utilization.

5. It was concluded that sensitizing sesquiterpene lactones and isothiocyanates could induce oxidative stress in mouse skin, possibly as a result of their reductive metabolism.     

A New Sesquiterpene Lactone from Tsoongiodendron Odorum Chun

Abstract

A new sesquiterpene lactone (1) was obtained from the cytotoxic fraction of 95% ethanol extract of root barks of Tsoongiodendron odorum Chun together with two known sesquiterpene lactones, costunolide (2) and parthenolide (3). The structure of 1 was elucidated as 5α, 6α, 7β, 10β-11α, 13-dihydro-4(15)-eudesmene-12, 6-olide on the basis of chemical and spectral evidence including X-ray diffraction analysis. Costunolide showed cytotoxic activity against human leukemia (HL-60) cell line. Parthenolide showed promising cytotoxic activities in vitro against HCT-8, Bel-7402, SKOV3, KB, HELA and EJ cell lines. Also, the cytotoxic ethyl acetate fraction of ethanol extract of the root barks from which three chemical components were isolated showed promising cytotoxic activities in vitro against KB, BGC-823, Bel-7402, HCT-8, HL-60 cell lines.