凡德他尼衍生物的抗肿瘤活性筛选研究

目的对合成的一系列凡德他尼衍生物进行体内外抗肿瘤活性的筛选,为寻找低毒高效的新型酪氨酸激酶抑制剂研究提供依据。方法体外筛选采用均相时间分辨荧光(HTRF)法和磺酰罗丹明B(SRB)法分别进行激酶和细胞的筛选;采用经典的急性毒性实验方法,并建立移植人非小细胞肺癌H1975裸鼠模型评价其抗肿瘤活性。结果 HTRF结果显示有6个活性较好的化合物(TY8115、TY8119、TY8122、TY8128、TY8129、TY8131),其中TY8115对VEGFR-2和EGFR抑制作用均好于凡德他尼;SRB结果显示这些活性化合物对选用的3种靶细胞(A431、H1975、A549)均有不同程度的抑制作用,其中TY8115的肿瘤细胞增殖抑制作用最明显,且对非靶细胞(MDA-MB-231)生长影响很小;急性毒性实验结果显示TY8115没有表现出毒性反应;体内抗肿瘤活性研究结果显示TY8115对肺癌H1975具有疗效,75、150 mg/kg TY8115对H1975的相对肿瘤增殖率分别为54.44%、39.54%。结论化合物TY8115具有良好的抗肿瘤活性,并且毒副作用小,具有发展成为一种新型酪氨酸激酶抑制剂的潜力。     

多靶标受体酪氨酸激酶抑制剂的合成与初步活性筛选

目的 以查尔酮为先导化合物,设计并合成多靶标受体酪氨酸激酶抑制剂。方法 查尔酮骨架上引入苯甲酰胺片段,设计、合成了35个不含共轭双键的查尔酮衍生物。采用ADP-GloTM方法评价化合物对EGFR, KDR和FGFR1 三种肿瘤相关RTKs的抑制活性;采用MTT方法评价化合物对 MCF-7、A549、K562的细胞增殖抑制活性。结果 大部分目标化合物具有较好的RTKs抑制活性和抗肿瘤活性,部分化合物的活性接近阳性对照的水平。结论 新型查尔酮衍生物具潜在抗肿瘤活性,为抗肿瘤药物的发现奠定了基础。     

阿魏酸衍生物的合成及其抗肿瘤活性研究

目的 设计、合成了一系列含有喹唑啉结构的阿魏酸衍生物，并对其抗肿瘤活性进行研究。方法 基于拼接原理，将阿魏酸与喹唑啉进行结合，并对喹唑啉6位进行修饰得到目标化合物，采用CCK-8法测试目标化合物对肺癌细胞A549的抗肿瘤活性，利用分子对接技术对目标化合物与表皮生长因子受体的结合模式进行模拟。结果 目标化合物的结构经HRMS(ESI)、¹H-NMR进行确证；体外抗肿瘤活性结果表明化合物11d、11f、12d和12g的IC₅₀低于阳性药吉非替尼，11c的IC₅₀值与吉非替尼相当。结论 化合物11d(IC₅₀=3.09 μmol·L^-1)对肺癌细胞A549抗肿瘤活性较强，值得深入研究。     

新型表皮生长因子受体抑制剂抗T790M型非小细胞肺癌作用

目的观察7个FCN系列表皮生长因子受体(EGFR)酪氨酸激酶抑制剂对H1975细胞体内外抗肿瘤作用。方法采用非小细胞肺癌(NSCLC)H1975细胞(T790M突变)和人表皮鳞癌A431细胞(WT-EGFR)体外增殖抑制试验对7个FCN系列化合物进行筛选,得到抗肿瘤活性强且选择性更好的化合物。通过细胞划痕实验、流式细胞术分别测定化合物对细胞迁移能力、细胞周期和凋亡的影响,通过H1975细胞裸鼠移植瘤实验明确化合物在体内对肿瘤生长的影响。结果 H1975细胞增殖实验筛选出高活性化合物FCN12、FCN14和FCN15,半数抑制浓度(IC50)分别为(103.33±12.10)、(115.17±7.69)和(128.63±32.72)nmol·L~(-1);经A431细胞增殖实验筛选出FCN12、FCN14,两者的体外抗细胞增殖抑制活性与对照药AZD9291相当(IC50>2μmol·L~(-1)),并可将H1975细胞阻滞于G1期;与对照药相比,FCN12、FCN14可明显增加细胞凋亡率(P<0.01)和抑制细胞的迁移(P<0.05),且呈浓度依赖性;体内移植瘤实验结果显示FCN12、FCN14可以明显抑制肿瘤的生长并缩小肿瘤体积(P<0.01)。结论新型EGFR抑制剂FCN12、FCN14对EGFR耐药型NSCLC具有显著的体内外抗肿瘤活性。     

吲哚-1,3,4-噁二唑类衍生物的合成及其体外抗肿瘤活性研究

目的 合成吲哚-1,3,4-噁二唑类衍生物,并进行体外抗肿瘤活性研究。方法 以吲哚-3-甲酰肼为起始原料,通过[4+1]环加成反应、水解反应、缩合反应得到目标化合物。采用MTT法测试目标化合物对HeLa、SCG-7901和MDA-MB-231细胞的体外抗肿瘤活性。采用克隆形成实验考察化合物6f对SCG-7901细胞增殖的影响。采用Annexin V-APC/PI双染法检测化合物6f对SCG-7901细胞凋亡和坏死的影响。采用DAPI染色法检测化合物6f对SCG-7901细胞凋亡核染色质形态学的影响。采用DCFH-DA染色法检测化合物6f对SCG-7901细胞中活性氧含量的影响。结果 合成了15个吲哚-1,3,4-噁二唑类衍生物,其结构经¹H-NMR、¹³C-NMR和HR-ESI-MS确证。部分化合物表现出良好的抗肿瘤活性,其中化合物6f对HeLa、SCG-7901和MDA-MB-231 3种肿瘤细胞株均表现出明显的抗增殖作用,且具有一定的选择性。进一步研究表明,化合物6f以浓度相关的方式促进细胞产生活性氧,抑制细胞增殖,诱导细胞凋亡。结论 部分吲哚-1,3,4-噁二唑类衍生物表现出良好的抗肿瘤活性,为该类化合物的进一步抗肿瘤活性研究提供思路。     

萘普替尼靶向表皮生长因子受体不同突变亚型的抗肿瘤作用研究

目的 研究萘普替尼对表皮生长因子受体（EGFR）不同突变亚型荷瘤小鼠肿瘤生长的影响。方法 建立野生型EGFR人表皮鳞癌A431、L858R/T790M双突变EGFR非小细胞肺癌H1975、Del19突变型EGFR非小细胞肺癌HCC827及HER2高表达的胃癌N87的裸鼠移植瘤模型,待肿瘤长至100 mm³左右将小鼠分组,A431荷瘤小鼠分组：对照组、阿法替尼12 mg/kg和萘普替尼3.50、1.75、0.87、0.29 mg/kg;H1975荷瘤小鼠分组：对照组、阿法替尼30 mg/kg和萘普替尼7.00、3.50、1.75 mg/kg;HCC827荷瘤小鼠分组：对照组、阿法替尼12 mg/kg和萘普替尼3.50、1.75、0.87、0.60、0.29 mg/kg;N87荷瘤小鼠分组：对照组、阿法替尼12 mg/kg和萘普替尼7.00、3.50、1.75、0.87、0.29 mg/kg,每组10只动物。分组后即开始ig给药,对照组给予同体积去离子水,A431、H1975、HCC827、N87荷瘤小鼠分别给药14、21、7、14 d。观察相对肿瘤体积（RTV）、肿瘤增殖率及对肿瘤质量的抑制率,在体考察萘普替尼的抗肿瘤作用。结果 A431荷瘤小鼠：与对照组比较,萘普替尼各剂量组RTV均显著减小（P<0.01）,肿瘤增殖率分别为26.6%、32.5%、34.8%、42.2%;瘤质量均显著降低（P<0.01）,抑瘤率范围在48.3%~73.9%;0.87 mg/kg为起效剂量,3.5 mg/kg剂量与阿法替尼12 mg/kg效果相当,3.5mg/kg组小鼠体质量显著降低（P<0.01）。H1975荷瘤小鼠：与对照组比较,萘普替尼各剂量组RTV均显著降低（P<0.05、0.01）,肿瘤增殖率分别为14.5%、38.2%、65.3%;3.5 mg/kg为起效剂量,与阿法替尼30 mg/kg作用相当;7.0、3.5 mg/kg组小鼠体质量显著降低（P<0.05、0.01）。HCC827荷瘤小鼠：与对照组比较,萘普替尼3.50、1.75、0.87、0.60 mg/kg组RTV降低极为显著（P<0.01）,肿瘤增殖率均小于40%;瘤质量呈不同程度地降低,抑瘤率范围52.9%~89.7%;0.6 mg/kg为起效剂量,1.75 mg/kg剂量与阿法替尼12 mg/kg作用相当,3.5、1.75 mg/kg组小鼠体质量显著降低（P<0.01）。N87荷瘤小鼠：萘普替尼7.00、3.50、1.75 mg/kg组RTV及瘤质量均显著低于对照组（P<0.05、0.01）,且肿瘤增殖率均小于40%,抑瘤率范围54.9%~95.00%;1.75 mg/kg为起效剂量,3.5 mg/kg与阿法替尼12 mg/kg作用相当;7.0、3.5 mg/kg组小鼠体质量显著降低（P<0.01）。结论 萘普替尼具有明显的抗肿瘤作用,其中对EGFR野生型或单突变型荷瘤抗肿瘤作用较好。关键词：萘普替尼;表皮生长因子受体;表皮生长因子受体酪氨酸激酶抑制剂;突变;非小细胞肺癌     

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。     

以c-Met为靶点的抗肿瘤系列化合物体内外药效筛选

目的　从8个LY系列肝细胞生长因子受体（c-Met）酪氨酸激酶抑制剂中筛选具有抗肿瘤活性的化合物,并进一步评价其体内外抗肿瘤作用。方法　首先采用均相时间分辨荧光技术（HTRF）对LY系列化合物进行初步筛选,观察它们对c-Met酪氨酸激酶的抑制作用;采用CCK8法观察筛选出的活性化合物在体外对人胃癌MKN-45、人神经胶质瘤U87MG、人肾癌Caki-1、人前列腺癌PC-3细胞株的增值抑制作用。建立人恶性胶质母细胞瘤U87MG裸小鼠移植瘤模型,考察活性化合物的抑瘤效果。结果　HTRF结果显示有4个活性较好的化合物（LY22,LY25,LY28,LY32）,其中LY28对c-Met抑制作用优于阳性对照药Crizotinib;CCK8结果显示这些活性化合物对选用的4种靶细胞均有不同程度的抑制作用,其中LY28对肿瘤细胞增殖抑制作用最明显;裸小鼠移植瘤实验显示,LY28可显著抑制U87MG裸小鼠移植瘤的增殖,40 mg/kg LY28抑瘤率达到78.13%。结论　化合物LY28具有较好抗肿瘤活性,具有进一步研发的价值。     

新型含嘧啶环吲唑衍生物的合成及初步生物活性研究

目的设计合成新型含嘧啶环的吲唑衍生物,并检测其对VEGFR-2酶的抑制活性。方法以3-甲基-6-硝基吲唑为起始原料,经N-甲基化、氢化还原、亲核取代、烷基化及亲核取代反应合成目标化合物;采用均相时间分辨荧光（HTRF）法测定目标化合物对VEGFR-2磷酸化的抑制作用。结果与结论合成了15个未见文献报道的新化合物,其结构经1H—NMR和MS谱确证。活性评价结果显示,该系列化合物对VEGFR-2酶均有抑制活性,其中化合物7a、7c、7i表现出较强的抑制活性,其抑制活性与阳性对照药帕唑帕尼接近,由此推测此类化合物可能具有潜在的抗肿瘤活性。     

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。     

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

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.