UPLC-TOF-MS法鉴定胀果甘草药渣中黄酮类成分

目的建立胀果甘草药渣中黄酮类成分的超高效液相色谱-高分辨飞行时间质谱(UPLC-TOF-MS)定性分析方法。方法 AgilentSB-C18柱(100mm×4.6mm,1.8μm);流动相乙腈-0.1%甲酸水溶液,梯度洗脱;体积流量0.4mL/min;柱温25℃;检测波长254nm。ESI离子源,飞行时间质谱检测器。对比自制对照品进行鉴别。结果共鉴定出8个黄酮类成分,分别为2’,4,4’-三羟基查耳酮、甘草查耳酮D、甘草查耳酮甲、4’-羟基-2’’,2’’-二甲基吡喃[5’’,6’’,6,7]黄酮、甘草黄酮C、光甘草酮、甘草黄酮B和kanzonolE。结论建立了一种简单、可靠的UPLC-TOF-MS方法对胀果甘草药渣中黄酮类成分进行了鉴定,对胀果甘草药渣综合利用有一定参考价值。     

新疆胀果甘草粗提物和精制物对宫颈癌细胞的影响

目的研究新疆胀果甘草(Glycyrrhiza inflata Batalin)粗提物和精制物体外抗宫颈癌的活性。方法利用超声辅助乙醇对新疆胀果甘草进行粗提取,得到粗提物后,进一步利用大孔树脂柱层析法对粗提物进行精制,得到精制物,并利用紫外分光光度法对粗提物及精制物总黄酮的含量进行测定,测得粗提物总黄酮含量为5.6%,精制物总黄酮含量为56.4%。将粗提物和精制物分别作为试药,顺铂为阳性药。采用人宫颈癌HeLa和SiHa细胞作为体外实验对象,用MTT法测定对宫颈癌细胞抑制率,用流式细胞仪测定胀果甘草精制物对人宫颈癌细胞的凋亡作用。结果新疆胀果甘草粗提物和精制物均能明显抑制宫颈癌HeLa和SiHa细胞的生长,且2种药的抑制效应呈时间和质量浓度依赖性,但在同一质量浓度下,精制物的抑制率更高。流式细胞测定结果显示,甘草粗提物、精制物能够显著诱导宫颈癌HeLa和SiHa细胞凋亡。结论新疆胀果甘草粗提物和精制物能够有效抑制宫颈癌细胞增殖,促进宫颈癌细胞凋亡,且精制物的抗癌活性明显优于粗提物。     

HPLC法测定新疆胀果甘草全草中甘草查尔酮A的含量

目的对新疆胀果甘草(Glycyrrhiza inflata Bat)全草中甘草查尔酮A(Lico A)进行含量测定。方法用HPLC法对胀果甘草根、茎和叶中Lico A的含量进行测定。色谱柱:依利特Hypersil ODS2C18(250mm×4.6mm,5μm)配预柱;流动相:甲醇-水-冰醋酸(60∶40∶1);流速:1.0mL·min~(-1);柱温:25℃;检测波长:372nm。结果 Lico A质量浓度在6.937 5~111μg·mL~(-1)范围内线性关系良好(r=0.999 9);平均加样回收率99.6%,RSD=1.5%;按此法测得的Lico A含量在根中为0.468%,茎中为0.254%,而在叶中未检出。结论甘草查尔酮A在甘草中主要分布在根和茎中,茎也可作为获取Lico A的新资源。     

新疆胀果甘草化学成分的分离与鉴定

目的研究中药胀果甘草(Glycyrrhiza inflataBatal.)生产甘草酸后的工业尾料的化学成分,为进一步合理利用这一传统中药资源提供依据。方法采用反复硅胶柱色谱、聚酰胺柱色谱、Sepha-dex LH-20凝胶柱色谱等方法进行分离纯化,并通过理化常数测定与光谱分析鉴定其化学结构。结果从胀果甘草药渣的体积分数95%乙醇提取物中分离鉴定了9个化合物,分别为甘草查耳酮A(licochalcone A,1)、2′,4,4′-三羟基查耳酮(2′,4,4′-trihydroxychalcone,2)、(-)α,2′,4,4′-四羟基二氢查耳酮((-)α,2′,4,4′-tetrahydroxydihydrochalcone,3)、甘草查耳酮C(licochalcone C,4)、甘草查耳酮D(licochalcone D,5)、刺甘草查耳酮(echinatin,6)、kanzonol E(7)、咖啡酸二十二酯(docosylcaffeate,8)、甘草次酸(glycyrrhetinic acid,9)。结论其中化合物2~5、7~9均为首次从甘草药渣中分离得到,化合物2、3、7、8均为首次从胀果甘草中分离得到。     

甘草查尔酮A对人脑胶质瘤U251细胞增殖和凋亡的影响

目的 探讨甘草查尔酮A对人脑胶质瘤U251细胞增殖和凋亡的影响.方法 将体外培养的人脑胶质瘤U251细胞分为四组(n=20),对照组予以等量0.9％NaCl处理,甘草查尔酮A低剂量组予以15 μmol/L甘草查尔酮A处理,甘草查尔酮A中剂量组予以30 μmol/L甘草查尔酮A处理,甘草查尔酮A高剂量组予以45 μmol...     

一种新型α-甲基查尔酮的制备、抗宫颈癌活性及对Akt-MDM2-p53信号通路的影响北大核心CSCD

课题组前期实验中发现,新疆胀果甘草中的微量特殊成分甘草查尔酮A(LicoA)具有较强的体外抗宫颈癌活性并对多种肿瘤干细胞标记物具有显著的下调作用,但是因其在胀果甘草中含量极低(约0.36%)^([1])并且合成制备成本昂贵,制约其开发利用。因此,本文旨在对LicoA进行结构修饰,合成一种具有替代LicoA潜力的新型查尔酮类化合物并研究其抗宫颈癌活性。     

胀果甘草化学成分的分离与鉴定

目的研究胀果甘草乙醇提取物的乙酸乙酯萃取部分,对其中的化学成分进行分离鉴定。方法采用硅胶、Sephadex LH-20柱色谱以及半制备型高效液相色谱等手段进行分离纯化,通过理化性质和各种波谱技术对化合物进行结构鉴定。结果从胀果甘草的乙酸乙酯萃取部分分离得到5个化合物,分别鉴定为刺芒柄花素(Formononetin,1)、异甘草素(Isoliquiritigenin,2)、达维荚蒾苷元(Dihydroisoliquiritigenin,3)、α-甘草次酸(α-Glycyrrhetic acid,4)和β-甘草次酸(β-Glycyrrhetic acid,5)。结论化合物3为首次从胀果甘草中分离得到。     

甘草查尔酮A抑制小鼠黑色素瘤B16F10细胞增殖机制研究

目的研究甘草查尔酮A抑制B16F10细胞增殖机制。方法 SRB法检测甘草查尔酮A对B16F10细胞增殖影响,Giemsa染色法观察细胞形态变化,比色法检测B16F10细胞内、外黑色素含量,Annexin V-FITC/PI双染检测细胞凋亡率,流式细胞术测定细胞周期分布,Q-PCR法检测细胞凋亡相关基因B淋巴细胞-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞周期蛋白(Cyclin E2)和细胞周期蛋白依赖性激酶-2(CDK2)的mRNA表达。结果甘草查尔酮A能有效抑制B16F10细胞增殖,呈现浓度依赖性和时间依赖性;随药物浓度增加,细胞增殖速度降低,细胞形态由树突状变为固缩圆球状,并伴有黑色素颗粒物出现,且细胞内、外黑色素含量呈浓度依赖性增加趋势,甘草查尔酮A能使细胞阻滞在G1期,在低浓度时,诱导细胞分化,高浓度时,诱导细胞凋亡;同时,甘草查尔酮A下调凋亡相关蛋白Bcl-2/Bax比率,抑制周期相关蛋白Cyclin E2、CDK2的mRNA表达。结论甘草查尔酮A抑制B16F10细胞增殖机制可能是通过使B16F10细胞G1期阻滞,进而诱导细胞分化和凋亡。     

异紫堇二酮体内外抗肿瘤活性研究

目的异紫堇二酮的体内外抗肿瘤活性及其作用机制的初步研究。方法通过异紫堇二酮对体外培养肿瘤细胞的生长抑制,体内荷S180瘤和荷H22瘤昆明种小鼠的肿瘤生长抑制、体重以及对小鼠免疫器官的影响(脾脏指数、胸腺指数),对其抗肿瘤活性进行综合评价,并通过流式细胞术检测异紫堇二酮作用下肺腺癌细胞(A549)的细胞凋亡及周期分布情况,初步探讨该化合物的抗肿瘤作用机制。结果异紫堇二酮对9种所选肿瘤细胞具有一定的生长抑制作用,IC50值在1.452×104～2.460×104 mol.L-1之间;流式细胞术检测结果显示异紫堇二酮可诱导A549细胞凋亡,使A549细胞阻滞于G0/G1期并随作用时间延长出现明显的凋亡峰;剂量为100和200 mg.kg-1.d-1时,异紫堇二酮对荷S180瘤和荷H22瘤小鼠的抑瘤率均大于40%,但剂量在200 mg.kg-1.d-1时,荷瘤小鼠的胸腺指数与脾脏指数相比较生理盐水组均有明显降低。结论异紫堇二酮具有一定的抗肿瘤活性,其主要是阻断肿瘤细胞由G0/G1期向S期转变来诱导肿瘤细胞死亡。     

甘草药渣化学成分的分离与鉴定

目的 对甘草药渣的化学成分进行研,为甘草资源的再利用提供依据。方法 利用硅胶、聚酰胺柱色谱及反复重结晶等方法进行分离纯化,根据理化性质及波谱分析对分离得到的化合物进行结构鉴定 。结果 分离得到 9 个已知化合物,分别鉴定为白桦酯酸（betulinic acid, 1）、光甘草酮(glabrone, 2)、甘草黄酮 C（licoflavone C, 3）、甘草黄酮 B（licoflavone B, 4）、3-羰基甘草次酸 （3-oxo-18β-glycyrrhetinic acid, 5）、芒柄花素（formonoetin, 6）、甘草黄酮 （licoflavone, 7）、β-谷甾醇 （β-sitosterol, 8）、胡萝卜苷 （daucosterol, 9）。结论 化合物 1~6 均为首次从甘草药渣中分离得到,化合物 1、4、5 为首次从胀果甘草中分离得到。     

Inhibitory effects of licochalcone A isolated from Glycyrrhiza inflata root on inflammatory ear edema and tumour promotion in mice

Licochalcone A, 3-a,a-dimethylallyl-4,4'-dihydroxy-6-methoxychalcone, from the root of Glycyrrhiza inflata Beta (Leguminosae) (Xin-jiang liquorice) showed anti-inflammatory action towards mouse ear edema induced by arachidonic acid (AA) and 12-O-tetradecanoylphorbol 13-acetate (TPA) by topical application. Anti-tumour promoting action of licochalcone A was also observed in vivo for mouse skin papilloma initiated by dimethylbenz[a]anthracene (DMBA) and promoted by TPA. It inhibited in vitro 32Pi-incorporation to phospholipids in HeLa cells promoted by TPA. A competitive interaction of licochalcone A with the TPA-receptors in the cell membrane has been suggested.     

Antispasmodic activity of licochalcone A, a species-specific ingredient of Glycyrrhiza inflata roots

Licochalcone A, a species-specific and characteristic retrochalcone ingredient of Glycyrrhiza inflata root, has been shown to possess multiple bioactive properties. However, its muscle relaxant activity has not been reported previously. Licochalcone A showed a concentration-dependent relaxant effect on the contraction induced by carbachol (50% effective concentration (EC50) = 5.64 +/- 1.61 microM), KCl (EC50 5.12 +/- 1.68 microM), BaCl2 (EC50 1.97 +/- 0.48 microM) and A23187 (EC50 2.63 +/- 2.05 microM). Pretreatment with licochalcone A enhanced the relaxant effect of forskolin, an adenylyl cyclase activator, on the contraction in a similar manner to 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase (PDE) inhibitor. Furthermore, the IC50 (22.1 +/- 10.9 microM) of licochalcone A against cAMP PDE was similar to that of IBMX (26.2 +/- 7.4 microM). These results indicated that licochalcone A may have been responsible for the relaxant activity of G. inflata root and acted through the inhibition of cAMP PDE.     

Synthesis and in vitro antitumor activity of novel ring D analogues of the marine pyridoacridine ascididemin: structure-activity relationship

Marine compounds with pyridoacridine skeletons are known to exhibit interesting antitumor activities. Ascididemin has already been reported as displaying significant antitumor activities in vitro and has also been found to have a relatively high global toxicity in vivo. We synthesized a series of 16 analogues (among which 11 compounds were different from previously described ones) with the aim of developing new anticancer agents with significantly improved efficacy/tolerability ratios. These compounds were obtained either by total synthesis from 5,8-quinolinedione and substituted 2-aminoacetophenones or by the direct substitution of ascididemin. The different compounds and ascididemin used as the control compound were tested at six different concentrations on 12 different human cancer cell lines of various histopathological types (glioblastomas and breast, colon, lung, prostate, and bladder cancers). The IC(50) value (i.e., the drug concentration inhibiting the mean growth value of the 12 cell lines by 50%) of these compounds ranged over five log concentrations, i.e., between 10 000 and 0.1 nM. For several new chemical entities, the antitumor activity (determined in vitro) and tolerability (determined in vivo) were superior to those of the parent alkaloids, i.e., ascididemin and 2-bromoleptoclinidone.     

六配位R2SnCl2（N—N）型新配合物的合成及其体外抑癌活性

从８０年代初起,有机锡化合物的抗癌活性日益受到重视,研究人员合成了大量的有机锡化合物,并对其进行了较全面的抗癌活性筛选〔１～３〕．吡唑衍生物具有抗菌〔４,５〕和杀虫作用〔６〕,其结构中有多个配位点,本文合成了两个新的吡唑衍生物（见图１）并用其作为双氮螯合配体来合成数个六配位Ｒ２ＳｎＣｌ２（Ｎ（Ｎ）型新配合物,并对它们的结构进行了表征,考察了它们对多种癌细胞的抑制作用,探讨了构效关系,结果表明这些六配位Ｒ２ＳｎＣｌ２（Ｎ（Ｎ）型新配合物主要对Ｐ３８８淋巴白血病有较强的抑制作用．配体的合成配体Ｉ〔Ｎ…     

Synthesis of new 1,3,4-benzotriazepin-5-one derivatives and their biological evaluation as antitumor agents

New derivatives of 1,3,4-benzotriazepin-5-one were designed and synthesized as structural analogues to the antitumor agents devazepide and asperlicin. An efficient and novel approach to the synthesis of 2-amino-1,3,4-benzotriazepin-5-one 2 was developed and its structure was confirmed. The newly synthesized derivatives were evaluated for their in vitro antitumor activity on 60 different cell lines. Compounds 8 and 9 displayed the most potent antitumor activity against several cell lines specifically ovarian cancer, renal cancer and prostate cancer, while compounds 5, 10 and 12 showed significant activities against UO-31 renal cancer cell line.     

1-苯胺基-5H-哒嗪并[4,5-b]吲哚类化合物的合成及体外抗肿瘤活性

目的设计并合成1-苯胺基-5H-哒嗪并[4,5-b]吲哚类化合物,评价其体外抗肿瘤活性。方法以5-乙酰氧基-6-溴-2-溴甲基-1-环丙基-1H-吲哚-3-羧酸乙酯为起始原料,经8～9步反应合成目标化合物;采用MTT法,测定了目标化合物对肿瘤细胞株Bel-7402和HT-1080的抑制活性。结果与结论合成了12个新化合物,其结构经1H-NMR和MS确证;多个化合物显示出良好的抗肿瘤活性,化合物10a和10d活性突出,对肿瘤细胞株Bel-7402和HT-1080的抑制活性分别是阳性对照药gefitinib的4倍和5倍,值得进一步研究。     

3-取代-5-氟-1-2-二氢-3H-吲哚-2-酮类化合物的合成及抗肿瘤活性

5-溴-1H-吲哚经氰基取代、Vilsmeier-Haack反应、水解、缩合制得3-[(Z)-(5-氟-1,2-二氢-2-氧代-3H-亚吲哚基)甲基]-1H-吲哚-5-甲酸,再和相应的胺类化合物反应制得10个3-取代-5-氟-1,2-二氢-3H-吲哚-2-酮类化合物.以舒尼替尼为阳性对照,用MTT法测试目标化合物对人乳腺上皮细胞HMEC的体外抑制活性,其中1c、1f、1g和1h在浓度为10 μmol/L时,对HMEC的抑制活性优丁舒尼替尼.进一步测试1c和1e对SGC7901、A549、HL-60、SK-BR-3、HCT116肿瘤细胞株的抗增殖活性.结果表明,1c和1e对白血病细胞株HL-60的抗增殖活性优丁舒尼替尼.     

Constituent properties of licorices derived from Glycyrrhiza uralensis, G. glabra, or G. inflata identified by genetic information

Constituent properties of licorices derived from Glycyrrhiza uralensis, G. glabra, and G. inflata are revealed by comparing 117 of licorice identified using four genetic markers; internal tracscribed spacer (ITS) on nuclear ribosomal DNA, rbcL gene, matK gene, and trnH-trnK1 intergenic region on chloroplast DNA. Regarding six main constituents of licorice; glycyrrhizin, liquiritin, liquiritin apioside, isoliquiritin, isoliquiritin apioside, and liquiritigenin, the constituent property of G. glabra resembles to that of G. inflata. On the other hand, the constituent property of G. uralensis is not similar to that of G. glabra or G. inflata and is characterized by a wide content variation of the six constituents compared to those of G. glabra and/or G. inflata. The mean contents of liquiritin, isoliquiritin, or liquilitigenin in G. uralensis are significantly higher than those of G. glabra or G. inflata. Therefore, the licorice species should be selected depending on these constituent properties for the traditional Chinese medicines or the Japanese Kampo medicines. Additionally, glycycoumarin, glabridin, and licochalcone A were reconfirmed as the species-specific typical constituents of G. uralensis, G. glabra, and G. inflata respectively. Therefore, it is resulted that the determination of the three species-specific constituents may be useful for the species identification of licorice. However, since 6% of licorice examined and hybrids were exceptions to the rule, their genetic information is necessary for the accurate species identification of licorice.     

氟喹诺酮C3杂环取代衍生物的合成及抗肿瘤活性研究(I)：环丙沙星噻二唑希夫碱

为发现氟喹诺酮类抗肿瘤先导化合物，用氨基噻二唑杂环替代环丙沙星(1)C3羧基形成环丙氟喹诺酮氨基噻二唑(2)骨架，然后与芳香醛缩合得到相应的希夫碱目标化合物(3a～3j)。新化合物的结构经元素分析和光谱数据表征，并用MTT法评价了它们体外对SMMC-7721、HL60和L1210 3种癌细胞株的生长抑制活性。结果表明，所合成的11个新化合物均具有潜在的体外细胞毒活性，其中目标化合物3d和3f的IC₅₀值达到微摩尔浓度数量级。这表明，氟喹诺酮类抗菌剂的3位羧基不是抗肿瘤活性所必需的，而被功能化修饰的杂环取代衍生物作为新结构抗肿瘤先导物具有进一步研究和开发的价值。