含有芳基脲结构的4H-吡喃类化合物的设计、合成及抗肿瘤活性研究

目的 设计并合成一系列含有芳基脲结构的4H-吡喃类化合物，评价该类化合物的体外抗肿瘤活性。方法 以间硝基苯甲醛、丙二腈和丙酮二羧酸二甲酯为原料，通过“一锅法”合成含有硝基的吡喃中间体，该中间体的硝基经铁粉还原为氨基，再与取代异氰酸苯酯反应得到一系列目标化合物。以人大细胞肺癌细胞H460、人肺癌细胞A549和人结肠癌细胞HT-29 3种肿瘤细胞为测试细胞株，采用MTT法评价了目标化合物的抗肿瘤活性。结果 合成了11个含有芳基脲结构的4H-吡喃类化合物。体外抗肿瘤活性试验表明，11个化合物对3种肿瘤细胞株均具有很好的抑制活性。其中化合物7c活性突出，对H460和A549细胞的IC₅₀值分别为0.82，0.98 μmol·L^-1, 优于阳性对照药索拉非尼(IC₅₀=3.20， 2.83 μmol·L^-1)。结论 含有芳基脲结构的4H-吡喃类化合物具有很好的抗肿瘤活性，可作为抗肿瘤化合物的结构骨架进一步研究。     

山胡椒内生真菌Trichoderma sp. SHJN1和Perenniporia sp. SHJG1的抗肿瘤活性成分研究

目的 从民族药山胡椒内生真菌Trichoderma sp.SHJN1和Perenniporia sp.SHJG1的代谢物中寻找活性先导化合物。方法 采用正相硅胶、反相硅胶、Sephadex LH-20凝胶及制备型HPLC等对Trichoderma sp.SHJN1和Perenniporia sp.SHJG1发酵物进行分离纯化，再通过NMR、ESI-MS等鉴定化合物结构，同时采用人乳腺癌细胞（MCF-7）和人肺癌细胞（A549）对这些化合物的抗肿瘤活性进行初步评价。结果 从2株内生真菌次级代谢产物中共分离鉴定了12个化合物：alantrypinone （1）、oryzalactam （2）、phomoindene A （3）、cis-gregatin B （4）、huaspenone B （5）、stigmasta-7，22-dien-3β，5α，6α-triol （6）、ergosterol （7）、1-deoxy-2-demethylviridiol （8）、viridiol （9）、trichodermamides A （10）、chromone （11）、对-羟基苯乙酸（12）。抗肿瘤活性评价结果显示，化合物3 抑制MCF-7细胞增殖活性IC₅₀为（62.9±1.02）μmol·L^-1[顺铂（cisplatin，DDP） IC₅₀为（30.1±1.67）μmol·L^-1]；化合物8 和9 抑制A549细胞增殖活性的IC₅₀分别为（34.6±1.57）μmol·L^-1和（44.9±1.74）μmol·L^-1[DDP IC₅₀为（20.6±1.42）μmol·L^-1]。结论 化合物3 ，8 和9 具有潜在抗肿瘤活性。     

二茂铁杂环类化合物的合成及抗三阴性乳腺癌活性研究

目的 设计、合成杂环二茂铁衍生物,并研究其抗三阴性乳腺癌活性。方法 以二茂铁查耳酮为先导化合物,对其进行结构改造,合成了一系列含有杂环的二茂铁衍生物,并通过CCK8试剂盒测试化合物抗乳腺癌活性。结果 合成了28个二茂铁衍生物,其结构均通过¹H-NMR和MS加以确证。初步的生物活性测试结果表明,所合成的二茂铁衍生物对三阴性乳腺癌MDA-MB-231细胞有较强的选择性和抑制活性,其中咪唑杂环化合物抗肿瘤活性强于相应的吡唑类和嘧啶化合物。尤其是28a[IC₅₀=（1.6±0.23）μmol·L^-1]对MDA-MB-231的抑制活性分别是先导化合物3[IC₅₀=（10.7±1.41）μmol·L^-1]和他莫昔芬[IC₅₀=（13.7±1.17）μmol·L^-1]的6和10倍,同时这些二茂铁衍生物对正常乳腺上皮细胞MCF-10A均没有毒性。结论 本研究为开发具有抗三阴性乳腺癌活性的化合物提供了信息和依据。     

白藤梨根体外抗肿瘤作用部位筛选及总三萜含量测定

目的 筛选白藤梨根体外抗肿瘤作用部位并测定总三萜含量。方法 采用RTCA DP实时细胞分析系统研究白藤梨根提取物对人肺癌A549细胞和人肝癌HepG2细胞体外增殖的抑制作用,筛选其抗肿瘤作用的活性部位;并通过紫外分光光度法测定白藤梨根活性部位中总三萜的含量。结果 白藤梨根各提取物对人肺癌A549细胞和人肝癌HepG2细胞均有一定的抑制作用,其中白藤梨根石油醚部位、乙酸乙酯部位和乙醇部位对人肺癌A549细胞的IC₅₀值分别为2,0.15,3.8 mg·mL^-1,对人肝癌HepG2细胞的IC₅₀值分别为75,7.6,71 μg·mL^-1;紫外分光光度法测得白藤梨根乙酸乙酯提取物中总三萜含量高达45%。结论 白藤梨根体外抗人肺癌细胞和人肝癌细胞的主要部位为乙酸乙酯部位,其中总三萜为其主要抗肿瘤成分。     

4(3H)-喹唑啉酮衍生物的合成及体外抗肿瘤活性

将6-溴甲基-2-甲基-4(3H)-喹唑啉酮在无水磷酸钾存在下与二硫化碳以及不同的胺反应，合成了一系列具有二硫代氨基甲酸酯侧链的4(3H)-喹唑啉酮衍生物，其结构经ESI-MS，¹H NMR，元素分析或HRMS所证实。采用MTT法测定了目标化合物8a~8q对人慢性髓性白血病K562细胞和人宫颈癌Hela细胞的体外抗肿瘤活性，结果表明化合物8q对K562和Hela细胞的体外生长具有显著的抑制作用，IC₅₀值分别为0.5和12.0 μmol·L^-1，因而可作为抗肿瘤药物研究的先导化合物。     

狼毒大戟中松香烷型二萜内酯的抗肿瘤活性研究

摘 要 目的： 对狼毒大戟中松香烷型二萜内酯类化学成分进行分离和结构鉴定,将得到的单体化合物与6种不同的人肿瘤细胞株相互作用探讨其抗肿瘤活性。方法: 通过硅胶、ODS等柱色谱与1D、2D-NMR波谱学测试技术结合,从狼毒大戟中分离并确定4个松香烷型二萜内酯类化合物：jolkinolide A (1)、jolkinolide B (2)、17 hydroxyjolkinolide A (3)、17-hydroxyjolkinolide B (4)。采用噻唑蓝比色法（MTT法）分别测定其与6种人肿瘤细胞株：肝癌HepG-2、乳腺癌MCF-7、胃癌SGC 7901、胃癌BGC-823、胃癌MGC-803、宫颈癌Hela相互作用结果,并计算抑制率与半抑制率(IC₅₀)。结果: 化合物2对肝癌HepG-2、乳腺癌MCF-7、宫颈癌Hela细胞的增殖有显著得抑制作用IC₅₀均＜25 μmol·L^-1,化合物4对乳腺癌MCF 7细胞的增殖有显著抑制作用IC₅₀＜30 μmol·L^-1。结论: 狼毒大戟中松香烷型二萜内酯类化合物具有显著的抗肿瘤活性。     

4-芳氨基-6-溴喹唑啉类化合物的合成及抗肿瘤活性的初步研究

目的 设计合成新的4-芳氨基-6-溴喹唑啉类化合物,并评价其抗肿瘤活性。 方法 以4-氯-6-溴喹唑啉和含有不同取代基的(E)-氨基二苯乙烯或4-[2-（2-呋喃基）]乙烯基苯胺为原料,经过亲核取代反应合成了5种目标化合物。采用MTT法,以人低分化胃癌细胞（BGC-823）和人肺癌细胞（A549）为受试细胞株对目标化合物进行体外抗肿瘤活性评价。 结果与结论 所得化合物的结构利用IR、1HNMR、13CNMR和元素分析进行了确认。5种化合物对BGC-823和A549两种细胞模型的体外抗肿瘤活性测试结果表明,大部分化合物具有较好的抗肿瘤活性。     

激光散射法测定吉非替尼原料药粒度

摘 要 目的：建立测定吉非替尼原料药的粒度及其分布的方法,并进行方法学验证。方法: 采用Malvern Mastersizer 3000 激光粒度分析仪和Hydro LV湿法进样器,以光散射法测定吉非替尼原料药的粒度分析并进行方法学考察,泵速为2 000 r·min^-1,遮光比为8%～20%,背景与样品的扫描时间为10 s,样品折射率为1.500,样品吸光率为0.00。结果: 3批吉非替尼原料药的D₁₀均小于7 μm,D₅₀均小于15 μm,D₉₀均小于25 μm,符合制剂终产品生产的需求。结论:该方法准确、简便、重复性好,适用于吉非替尼原料药的粒度分析。     

2,4-二氨基5-氟-6-取代苄氨基喹唑啉的合成及生物活性

本文报道2,4-二氨基-5-氟-6-取代苄氨基喹唑啉类化合物的合成及抗疟、抗肿瘤和抗菌活性,这类化合物的合成是由5-氟-2,4,6-三氨基喹唑啉(6a)与取代苯甲醛缩合成Schiff碱,然后经还原、甲酰化、亚硝化或甲基化制得。5-氟-2,4,6-三氨基喹唑啉(6a)尚未见文献报道,由5-氟-2,4-二氨基喹唑啉(4)经硝化生成异构体5a和5b分离得5a后再经还原制得。经对伯氏鼠疟原虫Plasmodium berghei抑制性治疗筛选,有6个化合物I_2,4.5,6和II_5,6以每日1mg·kg^-1,给药4天,抑制率为100%;体外抗肿瘤活性以I4最强,对L1210白血病细胞的IC₅₀为9.86×10^-4μg·mL^-1,优于氨甲蝶呤(MTX);经对18种常见菌进行体外筛选,发现对肺炎双球菌Diplococcus pneumoniae活性较好。     

甲氧基咔唑类化合物的合成及其体外抗肿瘤活性

目的设计合成新型甲氧基咔唑类抗肿瘤化合物。方法将4，5-二甲氧基-2-溴硝基苯与甲氧基碘代苯化合物在铜粉催化下经Ullmann反应制得甲氧基-硝基联苯化合物，再经亚磷酸三乙酯还原得三种甲氧基咔唑。对9-位氮进行修饰，合成了16个衍生物。利用核磁共振、质谱、红外光谱和元素分析进行了结构确认。结果体外初步抗肿瘤活性测试结果显示，目标物4_c，5_a，5_b，5_g，5_h，5_i，5_I，5n和5_p对人结肠癌HT-29细胞的IC₅₀值分别12.1，10.6，8.1，3.1，4.4，10.1及9.2 μmol·L^-1。目标物4_a对人口腔上皮癌KB细胞的IC₅₀值为17.7 μmol·L^-1。结论部分目标化合物对HT-29及KB肿瘤细胞具有较好的抑制作用。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.