中华真地鳖抗肿瘤蛋白纯化、鉴定及活性研究

目的分离纯化中华真地鳖(Eupolyphaga sinensis Walker)中抗肿瘤活性成分并研究其活性。方法采用硫酸铵沉淀、超滤、CM-Sepharose FF、DEAE-Sepharose FF及Q-Sepharose HP离子交换、Butyl Sepharose HP疏水色谱以及Superdex 75凝胶过滤色谱等技术分离纯化抗肿瘤组分;SDS-PAGE及MALDI-TOF质谱进行鉴定;采用MTT法考察其抗肿瘤活性。结果分离纯化得到一分子质量约为72 kD的抗肿瘤蛋白(EPS72)。该蛋白对肝癌Bel-7402细胞、肺癌A549细胞等多种人癌细胞株表现出较强的增殖抑制作用,并呈浓度依赖性。结论纯化的蛋白具有潜在的抗肿瘤作用。     

诺卡菌属菌株04-5195发酵液中活性成分04-5195A的分离、纯化与结构鉴定

目的分离、纯化与鉴定诺卡菌属菌株04-5195发酵液中的活性成分。方法采用多种层析方法进行活性成分分离，并通过FAB-MS、^1H-NMR和^13C-NMR测定对其结构进行鉴定。结果分离到一个主要组分04-5195A，其结构鉴定为3，4-二氢-8-羟基异香豆素衍生物。结论主要组分04-5195A与文献的报道的amicoumacin B相同。     

木蹄层孔菌石油醚组分的成分分析及抗肿瘤活性研究

目的:分析木蹄层孔菌(Fomes fomentarius)石油醚组分中甾醇和三萜类等成分及其主要抗肿瘤活性物质。方法:采用95%乙醇回流提取法及拌硅胶淋洗方法得到木蹄层孔菌的石油醚组分。用气相色谱-质谱联用技术对石油醚组分甾体和三萜类成分进行分析和鉴定并计算出其含量。用硅胶柱色谱法分离纯化含量最高的7,22-二烯麦角甾-3-酮。采用Alamar Blue法进行体外抗肿瘤活性检测。结果:从木蹄层孔菌石油醚组分中检测出37个成分,鉴定了其中的35个成分,包括10种甾体和三萜类成分。其主要成分为:7,22-二烯麦角甾-3-酮(37.67%)和5,24(28)-二烯麦角甾-3-醇(27.01%)。石油醚组分对细胞株NCI-H460的IC50为75μg.mL-1,7,22-二烯麦角甾-3-酮对细胞株NCI-H460的IC50为30μg.mL-1,抗肿瘤活性与单体浓度存在明显的量效关系。结论:木蹄层孔菌石油醚组分主要成分为7,22-二烯麦角甾-3-酮和5,24(28)-二烯麦角甾-3-醇,抗肿瘤活性物质最高的为7,22-二烯麦角甾-3-酮。研究结果为木蹄层孔菌的进一步开发提供了依据。     

蛇毒抗肿瘤蛋白化学成分的研究

对蛇毒抗肿瘤蛋白的化学成分进行研究。采用RP-HPLC分离蛇毒抗肿瘤蛋白各组分,收集纯化组分Ⅳ。非还原型SDS-PAGE和RP-HPLC鉴定组分Ⅳ的纯度;还原型SDS-PAGE与MALDI-TOF质谱仪测定组分Ⅳ的相对分子质量;测定组分ⅣN-末端氨基酸序列并进行Western blot鉴定;MTT法检测组分Ⅳ对体外培养的K562细胞的生长抑制作用。结果表明蛇毒抗肿瘤蛋白组分Ⅳ经SDS-PAGE和RP-HPLC分析为单一成分,MALDI-TOF质谱仪测得其相对分子质量为6714.22,其N-末端前5个氨基酸序列为L-K-C-N-K。经Western blot鉴定蛇毒抗肿瘤蛋白组分Ⅳ为细胞毒素,其对K562细胞的抑制作用呈明显的剂量依赖关系。     

皖南尖吻蝮蛇蛇毒中抗肿瘤活性蛋白分离纯化及其活体外性研究

目的:分离纯化皖南尖吻蝮蛇蛇毒(Wannan Agkistrodon acutus venom)中抗肿瘤活性蛋白并研究其活性。方法:利用DEAE-sepharose Fast Flow和SP-sepharose Fast Flow阴阳离子交换层析以及Sephadex G-75凝胶过滤层析等分离方法从皖南尖吻蝮蛇蛇毒中分离纯化得一种抗肿瘤活性蛋白,用CCK-8法检测该蛋白对体外培养的白血病K562细胞、结肠癌细胞(SW480)、胃癌细胞(SGC7901)、肝癌细胞(HepG2)的增殖抑制作用。结果:分离纯化得一相对分子质量约23700的抗肿瘤活性组分(ATF1-c),CCK-8检测对体外培养的K562、SW480、SGC7901、HepG2细胞的增殖抑制作用,呈剂量-时间依赖关系。结论:ATF1-c对体外培养的人癌细胞有明显的抑制和杀伤作用。     

猕猴桃根抗肿瘤作用研究

目的评价猕猴桃根提取物的体外、内抗肿瘤活性。方法猕猴桃根活性成分的分离、纯化采用传统的天然产物化学方法,猕猴桃根先用甲醇回流提取,然后用乙酸乙酯、氯仿、正丁醇萃取。不同组分对体外培养的肿瘤细胞增殖的作用采用磺酰罗丹明B方法;猕猴桃根的体内抗肿瘤作用采用小鼠肿瘤和人肿瘤裸小鼠移植瘤模型评价。结果氯仿提取物对肿瘤细胞增殖抑制作用最强,甲醇提取物次之。体内实验证实氯仿提取物有效地抑制小鼠肝癌模型和人肝癌裸小鼠移植瘤模型的生长,抑制率大概在38.0%。结论猕猴桃根提取物有一定的抗肿瘤作用;其活性成分主要存在于极性较小的组分。     

一种槲寄生多肽的一级结构分析和抗肿瘤活性

目的研究槲寄生枝叶中肽类抗肿瘤活性成分并阐明其结构。方法应用阳离子交换、凝胶过滤及高效液相等色谱方法进行分离纯化，基质辅助激光解吸飞行时间质谱用于测定质量数。采用Edman降解结合酶解法确定多肽的完整序列。对多肽用MTT法进行体外抗肿瘤活性评价。结果从槲寄生中纯化出一种新的多肽，命名为槲寄生毒素B2(viscotoxin B2)，其一级结构为KSCCKNTTGRNIYNTCRFAGGSRERCAKLSGCKIISASTCPSDYPK。Viscotoxin B2对体外培养的大鼠成骨样肉瘤细胞的IC₅₀为1.6 mg·L^-1。结论Viscotoxin B2与白果槲寄生中的槲寄生毒素有很高的同源性，并具有较强的抗肿瘤活性。     

白唇竹叶青蛇(Trimeresurus albolabris)毒降纤酶的分离纯化以及性质

目的从白唇竹叶青蛇(T.albolabris)毒中分离纯化无出血作用的降纤活性组分,探讨其理化性质及部分生物功能。方法用DEAE-SephadexA-25,SephadexG-100和CM-SephadexC-50三步色谱法进行分离纯化。SDS-PAGE和HPLC鉴定其纯度和相对分子质量,平板法测定其降纤活性。结果从白唇竹叶青蛇毒中分离纯化获得单一的降纤组分,能迅速水解纤维蛋白原或纤维蛋白原Aα链,缓慢水解Bβ链,而对γ链无作用,SDS-PAGE鉴定其相对分子质量为56000。EDTA能抑制其纤维蛋白原水解活性,而PMSF、β-巯基乙醇对其活性无影响,提示该组分为单链α金属蛋白酶。结论从白唇竹叶青蛇毒中分离纯化得到1种无出血作用且降纤活性强的新蛇毒降纤酶。     

链霉菌NOV-0827产生的抗肿瘤活性成分的分离纯化和结构鉴定

目的 分离、纯化和鉴定链霉菌NOV-0827发酵液中的抗肿瘤活性成分.方法 采用大孔吸附柱层析、硅胶柱层析、凝胶柱层析等方法对该菌株的次级代谢产物进行分离纯化;根据理化性质和波谱学方法进行化学结构的鉴定;利用MTT法来检测化合物的抗肿瘤活性.结果 从该菌株发酵液中共分离到3个化合物,NOV-0827-A、NOV-0827-B和NOV-0827-C,经波谱方法鉴定分别与文献中报道的氨基香豆素类(aminocoumarin)抗生素新生霉素,大环内酰胺类抗生素GT-32A和GT-32B同质.活性研究表明NOV-0827-B和NOV-0827-C均对结肠癌细胞SW620具有较强的增殖抑制活性,其IC50值分别1.107μmol/mL和5.358μmol/mL.结论 首次从同一陆生链霉菌菌株的发酵液中分离得到新生霉素及其他两个具有抗肿瘤活性的大环内酰胺类物质.     

海洋弧菌B2817的抗肿瘤活性成分研究

目的 对海洋弧茵B2817的抗肿瘤活性成分进行分离纯化和结构鉴定.方法采用溶剂萃取、制备HPLC等分离手段.对该菌株所产的活性成分进行了活性追踪分离.通过现代波谱学手段进行化学结构鏊定,以MTT法评价了化合物的抗肿瘤活性.结果从中分离得到4个化合物.分别鉴定为灵茵红素(1)、肉豆蔻酸(2)、7-十六碳烯酸(3)、7,10-十八碳二烯酸(4).化合物1对肿瘤细胞SM7721、S180具有强细胞毒活性.IC50分别为6.3、5.6μg·mL-1,而对人正常肝细胞HL-02无毒性.结论 4个化合物均为首次从该菌株中得到,而且弧菌B2817为灵茵红素的生产提供了新的来源.     

软骨抗癌活性成分抑制血管生成机理的研究

利用从牛软骨中提取的软骨抗肿瘤制剂、软骨血管生成抑制剂和软骨血管生成抑制因子，分别测定了它们对血管内皮细胞DNA合成、鸡胚绒毛尿囊膜血管生成以及内皮细胞迁移运动的抑制效应。提示软骨中提取的这些抗癌活性成分有可能为临床开辟一条以抑制肿瘤血管生成为特色的抗癌治疗新途径。     

Effect of linalool as a component of Humulus lupulus on doxorubicin-induced antitumor activity

As malignant neoplasm is a major public health problem, there is a need for the development of a novel modulator that enhances antitumor activity and reduces adverse reactions to antitumor agents. In this study, the effects of some volatile oil components in Humulus lupulus on doxorubicin (DOX) permeability in tumor cells and DOX-induced antitumor activity were examined. In vitro, DOX levels in tumor cells by combined linalool as its component significantly increased in the DOX influx system, and the increased effect by linalool on DOX cytotoxicity was shown. In vivo, the combination of DOX with linalool significantly decreased tumor weight compared with that of DOX alone treated group. The promotion of DOX influx level by combined linalool did not depend on energy, whereas it was suppressed by the absence of Na⁺. This promoting effect was suppressed by the presence of S-(4-nitrobenzyl)-6-thioinosine and inhibited dependently on phlorizin concentration. It is considered that linalool promoted DOX influx in tumor cells because of its action on DOX transport through concentrative Na⁺-dependent nucleoside transporter 3, which increased DOX concentration in tumor cells and thus enhanced the antitumor activity of DOX. Therefore, linalool as a food component is anticipated to be an effective DOX modulator.     

刺五加有效成分的抗肿瘤作用研究与评价

目的：研究刺五加有效成分的抗肿瘤作用进展。方法：分别介绍了刺五加中皂甙、多糖以及与黄芩联用的抗肿瘤作用研究,同时,对刺五加制剂治疗疾病的研究和评价做了总结,并对刺五加有效成分的抗肿瘤作用进行了展望。结果：刺五加有效成分具有一定的抗肿瘤作用。结论：临床医师应重视刺五加在抗肿瘤方面的作用,开发利用更多有效成分。     

Screening of host-mediated antitumor polysaccharides by crossed immunoelectrophoresis using fresh human serum

On crossed immunoelectrophoresis, human serum C3 (the third component of complement) converted by antitumor polysaccharides (ATSO [antitumor polysaccharide oral], AB-P [Agaricaus blazei polysaccharide], GU-P [Grifora umbellata polysaccharide], PS-K [polysaccharide Kureha] and zymosan) moved faster than native C3, appearing as the 3rd peak. The ratio of height of the 3rd peak to the alpha 2-macroglobulin (alpha 2-M) peak was linearly proportional to the dose of ATSO. At the dose of 500 micrograms/ml antitumor polysaccharides, the ratios were higher than 0.76, and the ratios for the serum treated with polysaccharide of no antitumor activity (dextran and gum arabic) were less than about 0.52. This ratio readily determined in vivo can be used as a measure for the antitumor activity of polysaccharides.     

糙皮侧耳糖蛋白的性质及体外抗肿瘤活性研究

目的：对糙皮侧耳真菌提取液进行纯化,并对纯化组分的理化性质及体外抗肿瘤作用进行研究。方法：用离子交换、凝胶过滤进行纯化,采用ＳＤＳ－ＰＡＧＥ测定蛋白分子量,用氨基酸自动分析仪测定氨基酸的组成,通过福林酚法测定其蛋白和多糖的含量,用薄层层析确定多糖的组成,将ＰＯＧＰ直接与Ｓ１８０腹水瘤细胞及小鼠正常细胞液混合,观察其活性。结果：纯化得到对肿瘤细胞具有细胞毒作用的糖蛋白（ＰＯＧＰ）,ＳＤＳ－ＰＡＧＥ显     

<Emphasis Type="Italic">Pulsatilla</Emphasis> saponin D: The antitumor principle from<Emphasis Type="Italic">Pulsatilla koreana</Emphasis>

By bioassay-guided separation, an already known saponin, Pulsatilla saponin D was isolated from the root of Pulsatilla koreana Nakai as a antitumor component when evaluated by in vivo antitumor activity as well as in vitro cytotoxic activity test. It showed potent inhibition rate of tumor growth (IR, 82%) at the dose of 6.4 mg/kg on the BDF1 mice bearing LLC cells.     

Role of the sugar moiety in the pharmacological activity of anthracyclines: development of a novel series of disaccharide analogs

The sugar moiety is an essential component of anthracycline antibiotics for their topoisomerase poisoning activity and antitumor efficacy. Since the sugar interacts with the minor groove, modifications in this moiety could enhance the recognition potential of the drug at the target level. Based on this hypothesis, novel anthracyclines, disaccharides lacking the amino group in the first (aglycone-linked) sugar, were designed. The 3'-amino group in the first sugar was replaced by an hydroxyl group, and the second sugar residue was bound to the first sugar via an alpha (1-4) linkage. The cytotoxic and antitumor activities of disaccharide analogs of idarubicin were critically dependent on the optimal (axial) orientation of the second sugar residue. Although configurational requirements of the sugar moiety for optimal drug activity support a critical role of the external (non-intercalating) drug domains in the interaction of anthracyclines with the DNA-topoisomerase (ternary complex), the antitumor efficacy of disaccharide analogs is not fully explained by effects mediated by the nuclear enzyme target. The development of this novel disaccharide series may provide insights for a rational synthesis of anthracycline analogs with improved pharmacological profile.     

54例新型抗肿瘤药物不良反应报告分析

目的:了解新型抗肿瘤药物不良反应的发生规律及特点,为临床合理用药提供参考.方法:收集2015年1月-2019年12月我院上报蚌埠市药品不良反应中心关于使用的新型抗肿瘤药物引起的54例不良反应进行分析.结果:54例新型抗肿瘤药物不良反应中,男性多于女性,以≥51岁的老年人群为主(45例,83.3％).涉及新型抗肿瘤药物1...     

Synthesis of biotinylated 18β-glycyrrhetinic acid and its effect on tumor cells activity

18β-Glycyrrhetinic acid (GA) is the active component in licorice root. Recent study has shown that GA exhibits many pharmacological activities. Here a new derivative of GA, biotinylated GA (BGA), was synthesized by introducing the biotin to the C-29 carboxyl of GA, and its antitumor effects were confirmed through tests on mouse B16 melanoma cells and BEL 7402 human hepatocarcinoma cells. At the same time, it is shown that the biotin group in BGA has no influence on the antitumor effects of GA. Then the stability of BGA in Roswell Park Memorial Institute (RPMI) medium 1640 and water was determined by high-performance liquid chromatography (HPLC), demonstrating much greater stability than GA in these two liquids.     

Predicting potential antitumor targets of Aconitum alkaloids by molecular docking and protein–ligand interaction fingerprint

Aconitine compounds are found in Aconitum Ouwu head, Chuan Wu, Aconitum, and other Ranunculaceae aconitum plants, which are not only active ingredient but also toxic component. In present study, 20 antitumor target proteins of different types were selected from Protein Data Bank (http://www.rcsb.org), for the purpose of finding potential antitumor targets of aconitum alkaloids, top ranked proteins were screened by molecular docking method, using the docking module in Sybyl-X 1.1 and Molecular Operating Environment (MOE) 2008, and screening result was verified by protein–ligand interaction fingerprint (PLIF) in MOE. Mesaconitine showed a C-shaped conformation when docking into heat-shock protein 90 (HSP90), which was similar with ANSA ring of geldanamycin. And the PLIF indicated that they shared many common amino acid residues interacted with HSP90; equally, Yunaconitine was found having similar conformation with the inhibitor of poly ADP-ribose polymerase-1 (PARP-1).