新型E-2,3-二芳基丙烯酰氧基膦酸酯衍生物的设计、合成与抗肿瘤活性

根据活性片段组合原理,设计、合成了一系列新型E-2,3-二芳基丙烯酰氧基膦酸酯衍生物,结构经IR、1H NMR、13C NMR及元素分析确证。采用MTT法测试目标化合物的抗肿瘤活性。结果表明:部分化合物对于所测试肿瘤细胞有抑制作用,其中化合物3e对A-549的活性[IC50=(12.7±1.9)μmol·L-1]最为突出,与对照药顺铂[IC50=(8.0±1.5)μmol·L-1]较为接近;化合物3g、3k对EC-109的增殖抑制作用最好,IC50分别为(9.5±1.8)μmol·L-1和(11.5±0.9)μmol·L-1;化合物3i、3k对SGC-7901、A-549、EC-109三种肿瘤细胞均有较好抑制作用。该类衍生物值得进一步研究。     

皂角刺抗肿瘤活性成分的分离鉴定与活性测定

目的分离鉴定皂角刺中抗肿瘤活性成分。方法用四甲基偶氮唑盐(MTT)比色法,结合硅胶、葡聚糖凝胶等色谱方法追踪分离皂角刺抗肿瘤活性成分,根据理化性质和波谱数据鉴定化学结构,用半数抑制浓度(IC50)评价其抗肿瘤活性。结果从皂角刺中分离得到8个化合物,分别为黄颜木素(1)、槲皮素(2)、3-βacetoxyolean-12-en-28-oic acid(3)、木栓酮(4)、棕榈酸(5)、白桦醇(6)、β-谷甾醇(7),胡萝卜苷(8)。化合物1、3对7种体外培养的肿瘤细胞抑制作用较好,在测定浓度范围内剂量依赖关系良好;化合物3对Bel-7402、HeLa、HT1080、KB、A549、SGC-7901、Heps等7种肿瘤细胞株生长增殖均具有良好的抑制作用,IC50为11.61~18.73 mg.L-1;化合物1对除A549之外的6种肿瘤细胞株抑制作用良好,IC50为11.34~19.32 mg.L-1。结论化合物2~6和8为首次从皂角刺中分离;化合物1、3的抗肿瘤活性为皂角刺中首次发现。     

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。     

橡胶籽种壳的化学成分研究

目的:研究橡胶籽种壳的化学成分。方法:通过LH-20和ODS-C18硅胶柱层析以及高压制备液相(HPLC)等色谱方法对橡胶籽种壳的化学成分进行分离,根据理化性质和光谱学数据鉴定化合物结构;采用MTT法评价分离产物对小鼠黑色素瘤细胞B16增殖的作用;应用人白细胞弹性蛋白酶(HLE)抑制剂高通量筛选模型评价分离产物的活性。结果:从橡胶种籽壳中得到9个化合物,分别鉴定为vanillin(1)、coniferal-dehyde(2)、erythro-guaiacylglycerol-β-coniferyl aldehyde ether(3)、threo-guaiacylglycerol-β-coniferyl aldehyde e-ther(4)、(-)-balanophonin(5)、isoamericanol A(6)、americanol A(7)、erythro-guaiacylglycerol-β-O-4’-de-hydrodisinapyl ether(8)、buddlenol A(9);化合物9对小鼠黑色素瘤细胞B16的增殖抑制作用较强,IC50值为20.6μmol.L-1;化合物5～9对人白细胞弹性蛋白酶有抑制活性,其IC50值分别为168,45.5,57.6,171和115μmol.L-1。结论:所有化合物均首次从该植物中得到,化合物9对B16肿瘤细胞的抗肿瘤活性以及化合物5～9的HLE抑制活性均首次报道。     

小叶锦鸡儿中异黄酮及其苷类化合物的体外抗肿瘤活性研究

目的研究小叶锦鸡儿中异黄酮及其苷类化合物的体外抗肿瘤细胞增殖作用。方法利用四唑盐比色法检测,观察小叶锦鸡儿中分离出的3种异黄酮及其苷类化合物:雁靛黄素(Ⅰ)、高丽槐树-7-O-β-D-吡喃葡萄糖苷(Ⅱ)和芒柄花素-7-O-β-D-吡喃葡萄糖苷(Ⅲ)对人乳腺癌细胞株、宫颈癌细胞株、人肝癌细胞株生长的影响。结果化合物Ⅰ对人乳腺癌细胞株、宫颈癌细胞株的增殖均有明显的抑制作用,IC50分别为20.2、16.9μg.mL-1;化合物Ⅱ对乳腺癌细胞株、肝癌细胞株均有不同强度的抑制作用,IC50分别为17.9、18.8μg.mL-1;化合物Ⅲ对乳腺癌细胞株的增殖有抑制作用,但不呈现浓度依赖关系,对其他两个细胞株的增殖不显示抑制活性。结论化合物Ⅰ、Ⅱ对两种细胞株有不同程度的抗肿瘤活性,化合物Ⅲ不具抗肿瘤活性。     

两种新硫色满酮衍生物体外抗肿瘤活性研究

目的研究新合成化合物（顺）-3-（氯代亚甲基）-5-氟-硫色满-4-酮（CFTK）和（顺）-3-（氯代亚甲基）-5-甲基-硫色满-4-酮（CMTK）对人乳腺癌MCF-7细胞、人肝癌HepG-2细胞、人结肠癌LS174T细胞、人肺癌A549细胞、人黑色素瘤A375细胞、人肾癌786-O细胞、人慢性粒细胞白血病K562细胞、人系髓样白血病U937细胞、小鼠肉瘤S180细胞的抗肿瘤活性,为深入研究CFTK和CMTK抗肿瘤作用提供实验依据。方法以不同浓度的CFTK和CMTK作用于9种癌细胞,应用改良的MTT法检测CFTK和CMTK对9种癌细胞生长的抑制作用。结果通过检测以上细胞的增殖情况,得出CFTK对9种癌细胞的半数抑制浓度（IC50）在（6.32±1.52）μmol.L-1~（19.8±4.08）μmol.L-1之间,CMTK对9种癌细胞的IC50在（7.16±0.76）μmol.L-1~（18.2±5.28）μmol.L-1之间;与同浓度的顺铂（CDDP）相比,具有明显的抗肿瘤活性。结论 CFTK和CMTK能够明显抑制肿瘤细胞的生长,具有极高的体外抗肿瘤活性,为进一步研究开发提供依据。     

2-苯氨基咪唑啉-5-酮衍生物的合成及抗肿瘤活性

目的设计并合成新型具有抗肿瘤活性的2-苯氨基咪唑啉-5-酮类化合物。方法以甘氨酸为起始原料,经环合、Knoevenagel缩合、N-烷基化、S-烷基化、取代5步反应合成2-苯氨基咪唑啉-5-酮类衍生物,并初步考察目标物对H460(人肺癌细胞)、HT29(人结肠癌细胞)以及A549(非小细胞肺癌细胞)的增殖抑制活性。结果与结论合成了11个未见文献报道的2-苯氨基咪唑啉-5-酮衍生物,化合物的结构经~1H-NMR、MS谱确证。体外活性测试结果显示,化合物1c、1e表现出较强的抑制细胞增殖活性,其对试验的3种肿瘤细胞的IC_(50)值分别为0.41、0.38(H460),0.41、0.66(HT29),1.44、0.75(A549)μmol·L~(-1),阳性对照药物顺铂(cisplatin)对3种肿瘤细胞的IC50值分别为0.22、0.62、0.35μmol·L~(-1)。初步探讨了目标化合物的构效关系,为进一步结构改造提供参考。     

4-叔丁基-5-(12,4,-三唑-1-基)-2-苄亚氨基噻唑的体外抗癌活性研究

目的研究4-叔丁基-5-(1,2,4-三唑-1-基)-2-苄亚氨基噻唑类衍生物对宫颈癌细胞系Hela、肝癌细胞系Bel 7402、鼻咽癌细胞系CNE 2的体外抗癌活性。方法细胞抑制率采用四甲基偶氮唑蓝(MTT)法测定。每种试样设置5个浓度梯度(0.025、0.05、0.10、.25、0.5μmol/ml),每个浓度取4个平行样,并设置无药对照实验,用半数抑制浓度(IC50)评价目标化合物的抗癌活性。结果被测化合物对3种癌细胞均表现出一定的抑制作用,其中化合物5对Hela癌细胞和Bel 7402癌细胞的IC50值分别为0.076 1、0.062 8μmol/ml;化合物6对CNE 2癌细胞的IC50值为0.050 8μmol/ml。结论生物活性测试表明该类型氨基噻唑衍生物具有较好的抗癌活性,值得进一步研究和关注。     

通光藤化学成分及抗肿瘤活性研究

目的 提取分离并筛选通光藤中具有抗肿瘤活性的化学成分,并研究其化学结构。方法 系统溶剂法对通光藤药材进行化学部位分离,并利用色谱法对正丁醇部位中的化学成分进行分离和纯化,利用波谱学方法鉴定其化学结构。MTT法筛选所得化学成分体外对8种人癌细胞增殖的抑制作用。结果 在通光藤正丁醇部位中得到3个化学成分单体分别为：牛奶菜醇（化合物1）、通光藤苷元B（化合物2）、大叶牛奶菜苷丁（化合物3）,化合物2对于MGC-803肿瘤细胞抑制作用较强,IC50为80.3μg.mL 1,而化合物3对于ISMMC-7721肿瘤细胞液表现出较强抑制作用,IC50为40.7μg.mL 1。结论 化合物2为首次在该植物中分离到的苷元成分。化合物2、3体外对肿瘤细胞具有显著的抑制活性。     

补骨脂定对骨髓间充质干细胞向癌相关成纤维细胞分化的抑制作用

目的探讨不同来源的间充质干细胞(MSCs)在癌细胞诱导下分化成癌相关成纤维细胞(CAFs)的情况,观察补骨脂定对人骨髓间充质干细胞(h BMSCs)和人脐带间充质干细胞(h UCMSCs)向CAFs分化的影响。方法利用Transwell小室,建立h BMSCs和h UCMSCs与4种女性特发恶性肿瘤细胞(BT-549、HEC-1B、Hela和SK-OV-3)共培养模型。梯度浓度的补骨脂定处理h BMSCs和h UCMSCs后,用Compu Syn软件计算半数抑制浓度(IC50),后续用10μmol·L-1补骨脂定处理各组细胞。用细胞增殖毒性(CCK-8法)检测细胞的增殖活性;流式细胞术分析细胞中α-平滑肌肌动蛋白(α-SMA)表达水平;用免疫印迹检测细胞中p-JAK2和p-STAT3表达水平。结果补骨脂定对h BMSCs和h UCMSCs的IC50分别是477.34和364.83μmol·L-1;在10μmol·L-1浓度下,对两种细胞活性的抑制率分别为(3.77±1.65)%和(3.70±2.11)%,均小于5%。h BMSCs与4种癌细胞共培养后,h BMSCs中α-SMA表达阳性率上升,肿瘤细胞的增殖活性增强,其p-JAK2和p-STAT3的表达水平上升;补骨脂定能显著抑制共培养体系中的上述现象。在共培养体系中,h UCMSCs中α-SMA表达阳性率不变,癌细胞的增殖活性却受抑制,其p-JAK2和p-STAT3的表达下降;补骨脂定不影响h UCMSCs中α-SMA、p-JAK2和p-STAT3的表达,但能增强h UCMSCs对癌细胞活性的抑制作用。结论在Transwell共培养环境中,补骨脂定可抑制h BMSCs向CAFs分化,降低癌细胞恶性增殖活性。     

金银花总黄酮对氧化应激中肝星状细胞的保护作用

目的:研究金银花总黄酮对氧化应激中肝星状细胞(HSC)的保护作用。方法:培养大鼠HSC为模型。实验分为空白对照(DMEM培养基)组、模型(DMEM培养基)组与金银花总黄酮1、2、3、4(12.5、25.0、50.0、100.0μg/ml)组。检测MTT抑制率;测定超氧化物歧化酶(SOD)与乳酸脱氢酶(LDH)活性、丙二醛(MDA)含量、总抗氧化能力(T-AOC)。结果:与空白对照组比较,模型组MTT抑制率降低,MDA含量增加,LDH活性增强,SOD活性减弱,T-AOC减弱,差异有统计学意义(P<0.01或P<0.05);与模型组比较,金银花总黄酮1、2、3、4组MTT抑制率升高,MDA含量减少,LDH活性减弱,SOD活性增强,T-AOC增强,差异有统计学意义(P<0.01或P<0.05)。结论:金银花总黄酮具有抑制肝纤维化形成的作用,其作用机制可能与抑制HSC的增殖及抗氧化应激,以及抑制脂质过氧化反应有关。     

Flavonoids and tyrosine nitration: structure-activity relationship correlation with enthalpy of formation.

The ability of 11 flavonoids, naturally occurring polyphenols, and their related structure-activity relationships (SAR's) for inhibiting peroxynitrite-induced nitration of tyrosine was investigated. The flavonoids under study could be classified into four groups having very distinct in vitro inhibition effects. We also calculated the heat of formation (DeltaH(f)) of the corresponding flavonoids radicals which supported this finding. The most effective flavonoids included: catechin, taxifolin, luteolin, quercetin, and myricetin which have a common structural feature of ortho-dihydroxyl moiety (3',4'-OH substitution). Naringenin, kaempferol, and morin were 50% less effective inhibitors than the former group of flavonoid while their activities were in the range of trolox (an alpha-tocopherol analogue). The common structural aspect of this group of flavonoids is 4'-OH substitution. Therefore, these two groups of flavonoids may have similar mechanisms for their inhibition activity. No inhibition activity was observed by galangin. Apigenin behaved as a pro-oxidant in our in vitro study. Naringin was as effective as the second group at 4 mM tyrosine concentration while did not illustrate any inhibitory effect at 1 mM concentration of tyrosine. Our study provides further evidence for the importance of the catechol B ring and to a lesser effect the importance of 4'-OH substitution. Moreover, we observed very little or no influence on activity of flavonoids by 3-OH substitution and/or a C2-C3 double bond conjugated with 4-keto group within the subgroup containing the catechol moiety. Theoretical calculation of DeltaDeltaH(f) for tyrosyl radical repair by flavonoids (TyO*+FlOH-->TyOH+FlO*) correlated well with our in vitro results (inhibition% = -10 (DeltaDeltaH(f)), R2=0.906). Furthermore, this correlation was independent of tyrosine concentration. This model can be used to accurately predict the inhibitory effect of flavonoids on nitrotyrosine formation.     

藤茶总黄酮体外抗肿瘤实验研究

目的观察藤茶总黄酮在体外对肿瘤的影响作用。方法应用四甲基偶氮唑蓝法研究藤茶总黄酮对肿瘤细胞体外增殖的影响。结果藤茶总黄酮对人乳腺癌细胞MCF-7、人前列腺癌细胞PC3和人前列腺癌细胞LNcap的抑制作用都比较强,且剂量依赖性比较明显,随着药物浓度的增大抑制率也随之增大。72 h半数抑制浓度IC50分别为19.15、163.98、27.54μg/mL。结论藤茶总黄酮在体外对人乳腺癌和人前列腺癌有明显的抑制作用。     

Inhibition of pro-inflammatory markers in primary bone marrow-derived mouse macrophages by naturally occurring flavonoids: analysis of the structure-activity relationship

Flavonoids possess several biological/pharmacological activities including anticancer, antimicrobial, antiviral, anti-inflammatory, immunomodulatory and antioxidant. The aim of this study was to evaluate the effect of flavonoids on macrophage physiology. For this purpose we selected some flavonoids belonging to the most common and abundant groups (flavonols--quercetin and kaempferol; flavones--diosmetin, apigenin, chrysin and luteolin; isoflavones--genistein and daidzein and flavanones--hesperetin). We decided to use primary bone marrow-derived macrophages (BMDM) as cellular model, since they represent a homogenous, non-transformed population of macrophages that can be stimulated in vitro to proliferate by macrophage colony-stimulating factor (M-CSF) or activated by LPS. In this regard, we demonstrated that most of the flavonoids assayed reduce macrophage M-CSF-induced proliferation without affecting cellular viability. Moreover, some flavonoids also inhibit TNFalpha production as well as iNOS expression and NO production in LPS-activated macrophages, an effect that has been associated with the inhibition of the NF-kappaB pathway. We also found that luteolin and quercetin are able to stimulate the expression of the anti-inflammatory cytokine IL-10 at low concentrations (<50microM). Analysis of the structure-activity relationship showed that four hydroxylations at positions 5, 7, 3' and 4', together with the double bond at C(2)-C(3) and the position of the B ring at 2, seem to be necessary for the highest anti-inflammatory effect.     

Structure-activity studies of flavonoids as inhibitors of hyaluronidase

The order of decreasing potency for five most potent flavonoids as inhibitors of hyaluronidase was found to be: condensed tannin less than luteolin less than apigenin less than kaempferol less than silybin. Kinetic studies of these inhibitors showed that their mode of inhibition was competitive. Aglycones were stronger inhibitors than their corresponding glycosides. The following flavonoid structure conferred potent inhibitory effect: a double bond between carbons 2 and 3; unsubstituted hydroxyl groups at positions 5, 7 and 4' and a ketone group at position 4.     

姜黄素抑制子宫颈癌HeLa细胞的作用

目的：探讨姜黄素抑制子宫颈癌HeLa细胞的作用及其机制。方法以细胞培养，镜下观察细胞形态学改变和计数法测定生长曲线；用3H-脱氧胸苷掺入法测定对DNA合成的影响，以MTT比色法检测给药后HeLa细胞的增殖抑制情况。结果姜黄素作用HeLa细胞后，癌细胞生长延缓并萎缩，胞质粗糙，有大量颗粒状物堆积，而且药物浓度越大，形态学改变越明显；生长曲线测定、3H-脱氧胸苷掺入法及MTT比色法实验结果显示，姜黄素对HeLa细胞的增殖和生长有显著的抑制作用并呈明显的时间一剂量依赖关系，当姜黄素浓度为20μg／ml以上时，其对HeLa细胞的掺入抑制率高于5一Fu20μg／ml对该细胞的掺入抑制率。结论姜黄素对HeLa细胞具有直接杀伤作用，其机制可能通过干扰细胞代谢，改变细胞外暌的性质抑制肿瘤细胞增值。     

乌骨藤提取物抑制慢性髓性白血病（K562）细胞增殖

目的提取乌骨藤中抗肿瘤成分,探讨其对慢性髓性白血病K562细胞增殖的抑制作用及机制。方法设计乌骨藤提取物浓度梯度、时间梯度干扰K562细胞增殖,通过显微镜观察细胞形态、MTT法检测增殖抑制率。结果随着浓度和时间梯度的递增,K562细胞增殖速度及细胞形态受到了明显的影响。结论乌骨藤提取物对慢性髓性白血病（K562）细胞增殖有明显的抑制作用。     

Inhibition of lipopolysaccharide-induced nitric oxide production by flavonoids in RAW264.7 macrophages involves heme oxygenase-1

The role of heme oxygenase-1 (HO-1) played in the inhibitory mechanism of flavonoids in lipopolysaccharide (LPS)-induced responses remained unresolved. In the present study, flavonoids, including 3-OH flavone, baicalein, kaempferol, and quercetin, induced HO-1 gene expression at the protein and mRNA levels in the presence or absence of LPS in RAW264.7 macrophages. This effect was associated with suppression of LPS-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) protein expression. Hemin induced HO-1 protein expression and this was associated with the suppression of LPS-induced NO production and iNOS protein expression in a dose-dependent manner. In addition, an increase in bilirubin production was found in flavonoid- and hemin-treated cells. Hemin, at the doses of 10, 20, and 50 microM, dose-dependently stimulated the flavonoid (50 microM)-induced HO-1 protein expression, and enhanced their inhibitory effects on LPS-induced NO production and iNOS protein expression. Pretreatment of the HO-1 inhibitor, tin protoporphyrin (10 microM), attenuated the inhibitory activities of the indicated flavonoids on LPS-induced NO production. Morphologic analysis showed that 3-OH flavone, baicalein, kaempferol, quercetin, hemin, and tin protoporphyrin did not cause any change in cell viability in the presence or absence of LPS. In contrast, only 3-OH flavone showed a significant inhibition of cell growth using the MTT assay. Transfection of an HO-1 vector in macrophages (HO-1/RAW264.7) resulted in a 3-fold increase in HO-1 protein compared with that the parental RAW264.7 cells. NO production mediated by LPS in HO-1 over-expressed RAW264.7 cells (HO-1/RAW264.7) was significant less than that in parental RAW264.7 cells. 3-OH Flavone, baicalein, kaempferol, and quercetin showed a more significant inhibition on LPS-induced NO production in HO-1/RAW264.7 cells than in parental RAW264.7 cells. These results provide evidence on the role of HO-1 in the inhibition of LPS-induced NO production by flavonoids. A combination of HO-1 inducers (i.e. hemin) and flavonoids might be an effective strategy for the suppression of LPS-induced NO production.     

通过碳碳双键连接的吲哚-2-酮与喹唑啉-4（3H）-酮杂合物的合成与抗肿瘤活性评价

通过2-甲基4-氧代喹唑啉-6-甲醛与不同的吲哚-2-酮的缩合反应,制备了一系列吲哚-2-酮与喹唑啉-4（3H）-酮的杂合物5a-j。MTT法测试结果表明,只有化合物5e对人肿瘤细胞A549、MCF-7、HeLa、HT-29和HCT-116表现出一定的细胞毒活性,50μM浓度下的抑制率为32．0％-62．3％。     

Effects of flavonoids on cyclic AMP phosphodiesterase and lipid mobilization in rat adipocytes.

Thirty-one flavonoids were tested for their effects on low Km phosphodiesterase with cyclic AMP as the substrate. Quercetin, luteolin, scutellarein, phloretin and genistein showed inhibitory potencies comparable to or greater than 3-isobutyl-2-methylxanthine (EC50 30-50 microM). Only four compounds namely, catechin, epicatechin, taxifolin and fustin stimulated the enzyme activity (stimulatory EC50 130-240 microM). The most potent phosphodiesterase (PDE) inhibitors were aglycones that had a C2.3 double bond, a keto group at C4 and hydroxyls at C3' and/or C4'. However, when the C-ring is opened then the requirement for the C2.3 double bond is eliminated. The same series of flavonoids were also tested for their lipolytic activity. The structural features required for effective synergistic lipolysis (with epinephrine) were generally similar to that required for potent PDE inhibition except that, for lipolytic activity, an intact C-ring was necessary. Fisetin and quercetin having the above-mentioned structure showed a dose- and time-dependent increase in lipolysis which was synergistic with epinephrine. Only butein and hesperetin showed inhibition of epinephrine-induced lipolysis, and their effect was dose-dependent. A time-course study indicated that hesperetin was able to delay the lipolytic action of epinephrine. It is most likely that the lipolytic effects of these compounds were not a result of PDE inhibition, as the orders of potency for the two activities had poor correlation. Apparently, the effective lipolytic flavonoids were also potent PDE inhibitors but not all the PDE inhibitors were able to induce lipolysis.