CDAP活化多糖制备1型肺炎链球菌荚膜多糖-蛋白结合物原液的稳定性评价

目的 评价以1-氰基-4-二甲氨基-吡啶四氟硼酸盐(1-cyano-4-dimethylamino pyridinium tetrafluoroborate,CDAP)活化多糖制备的1型肺炎链球菌荚膜多糖-破伤风类毒素结合物原液的稳定性.方法 以CDAP作为活化剂,连续制备3批多糖-蛋白结合物原液,分别放置于(37±2)℃和2～8℃,定期取样检测,并评价其稳定性.结果 多糖-蛋白结合物原液在(37±2)℃条件下保存21 d,2～8℃条件下保存12个月,其主要检测指标均符合质量要求,游离多糖和游离蛋白含量分别低于30.0％和5.0％,分配系数≤0.35的多糖回收率均＞65％.结论 确定了不同条件下1型肺炎链球菌荚膜多糖-破伤风类毒素结合物原液的有效保存时间.     

HPLC-ESI/ITMSn法分析大鼠体内4-甲基哌嗪-1-二硫代甲酸-(3-氰基-3，3-二苯基)丙酯盐酸盐及其主要代谢物

目的研究新化合物4-甲基哌嗪-1-二硫代甲酸-(3-氰基-3，3-二苯基)丙酯盐酸盐(TM208)在大鼠体内的主要代谢产物。方法大鼠ig TM208 500 mg·kg^-1后，收集粪样、尿样和血样，用液相色谱-电喷雾离子阱质谱法测定。根据TM208及其代谢产物的色谱保留时间和电喷雾离子阱质谱(ESI-ITMSⁿ )电离规律及生物体内药物代谢转化规律，推导代谢物的结构。结果在粪样中发现8种I相代谢产物，在尿样和血样中发现5种I相代谢产物，未发现II相代谢产物。结论本法操作简便、快速、灵敏度高、专属性强，是一种研究TM208体内代谢产物的有效方法。     

Studies on the metabolism of 4-methyl-piperazine-1-carbodithioc acid 3-cyano-3,3-diphenylpropyl ester hydrochloride in rats by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry

4-Methyl-piperazine-1-carbodithioc acid 3-cyano-3,3-diphenylpropyl ester hydrochloride(TM208) is a newly synthesized compound, which has shown excellent in vivo and in vitro anticancer activity and low toxicity. In this study, the metabolism of TM208 in rats was studied for the first time by high-performance liquid chromatography coupled with tandem mass spectrometry. Following a single oral administration to rats, TM208 was metabolized to eight metabolites (M1–M8). M1 is the desmethyl metabolite and the acylation of M1 with N-acetyl transferase results in M6 (N-acetyl metabolite), M5 is N-formyl metabolite; M4 is phenyl monohydroxylation metabolite, M2 is the sulfine metabolite of TM208, and M3 is also an odd-oxygen added products which the possible oxidation site has described in this paper; M8 is the metabolite resulting from the replacement of ‘–CS’ with ‘–CO’, M7 is a ring-opened piperazine oxidation products to a kind of acid.     

Valerian-Derived Sedative Agents. I. On the Structure and Spectral Assignment of the Constituents of Valmane Using the Selective INEPT Nuclear Magnetic Resonance Technique

The valepotriates, a group of chemically unstable iridoid triesters possessing sedative activity, contain various ester groups at the C-l, C-7, and C-11 positions. Using the selective INEPT NMR technique and employing a suitable polarization delay for long-range coupling, it was possible to achieve the assignment and location of the ester groups directly, without ambiguity, and without chemical modification. Six valepotriates isolated from Valmane tablets served as examples to demonstrate the utility of this NMR technique. During the course of this work, the acevaltrate fraction was shown to be a mixture of 1--ace valtrate (3) and 7--acevaltrate (4), the structures of valtrate (1) and didrovaltrate (2) were confirmed directly, and two new valepotriates, 5a and 5b, were obtained as an inseparable mixture and characterized.     

人C1酯酶抑制剂初步纯化过程pH值和聚乙二醇含量的优化#br#

目的 探讨pH值、聚乙二醇（polyethylene glycol,PEG）含量和离心条件对PEG4000粉末用于去除人C1酯酶抑制剂（C1 esterase inhibitor,C1-INH）制备原料中的IgM效果的影响,并通过对羧甲基（carboxymethyl,CM）离子交换层析中洗脱盐离子浓度的筛选,分离活性与非活性C1-INH。方法 向不同pH值的C1-INH制备原料中加入不同质量分数的PEG4000,于不同条件下离心后,用特定蛋白检测仪对离心后上清液中IgM和C1-INH含量进行检测,确定PEG沉淀法纯化C1-INH的最佳条件。将离心后上清液调节pH后作为CM离子交换层析上样样品,使用不同盐离子浓度的洗脱液对活性C1-INH进行分离,确定最佳的盐离子浓度。结果 在弱酸性pH（6.8）下,当PEG4000质量分数为12%,离心条件15 000×g、25 ℃、20 min时,IgM去除率>99%,且C1-INH的回收率>80%;在盐离子浓度为200 mmol/L时,产物中C1-INH的比活性最大（4.43 IU/mg）,且绝大多数杂质蛋白得以去除。结论  优化条件下PEG4000能有效去除C1-INH制备原料中的IgM,且能保持较高的C1-INH回收率;CM离子交换层析能对活性与非活性C1-INH进行有效分离,并去除大多数杂质蛋白。     

Modulation of the acceptance of transfer ribonucleic acids for amino acids by alkyl-aryltriazenes and imidazoles

Selected triazene and imidazole compounds were incubated with postmitochondrial supernatants from rat liver, and tRNA isolated from these incubates was charged with thirteen different amino acids. The majority of tested compounds enhanced the acceptance of initiator tRNA but inhibited the formation of l-leucyl-tRNA. No unequivocal results were obtained with the other tRNA species. The direct alkylating monomethyltriazenes enhanced the acceptance of initiator tRNA when incubated alone with unfractionated tRNA whereas the procarcinogenic dialkyltriazenes showed this effect only after preincubation with postmitochondrial supernatant containing the activating enzymes. It appears that the reactive molecular species, that arise from monomethyltriazenes by heterolysis, or from dimethyltriazenes by enzymic activation, modify the structure of tRNA in a specific manner, possibly by a chemical reaction. The enhanced acceptance of initiator tRNA seems to be indicative of the generation of electrophilic (potentially carcinogenic) intermediates from the incubated compounds.     

SDF—1a／54／KDEL基因电穿孔Molt-4细胞株的转染条件优化

目的探索以绿色荧光蛋白（GFP）为报告基因的重组质粒pEGFP／SDF-1a／54／KDEL电穿孔悬浮培养的Molt-4细胞的高效转染条件。方法通过控制脉冲幅度、脉冲电压、细胞生长状态等三个主要条件,采用不同条件组合后用电穿孔法将转入悬浮培养的Molt-4,用荧光显微镜和流式细胞仪观察转染率。结果传代后第二天的Molt-4细胞在260V、950μF得到最大转染率51．2％。荧光显微镜下可观察到被转染目的基因的细胞发出绿色荧光。结论优选260V、950μF和良好的细胞生长状态等条件下的Molt-4,可获得目的基因SDF—1a／54／KDEL的最高转染效率。     

Design and synthesis of leucine‐linked quinazoline‐4(3H)‐one‐sulphonamide molecules distorting malarial reductase activity in the folate pathway

Optimization of a modified Grimmel's method for N‐heterocyclization of a leucine‐linked sulfonamide side‐arm at position 2 leading to 2,3‐disustituted‐4‐quinazolin‐(3H)‐ones was accomplished. Further, 22 hybrid quinazolinone motifs ( 4a‐v ) were synthesized by N‐heterocyclization reaction under microwave irradiation using the ionic liquid [Bmim][BF₄]‐H₂O as green solvent as well as the catalyst. The in vitro screening of the hybrid entities against the malarial species Plasmodium falciparum yielded five potent molecules 4l, 4n, 4o, 4t , and 4u owning antimalarial activity comparable to those of the reference drugs. In continuation, an in silico study was carried out to obtain a pharmacophoric model and quantitative structure–activity relationship. We also built a 3D‐QSAR model to procure more information that could be applied to design new molecules with more potent Pf‐DHFR inhibitory activity. The designed pharmacophore was recognized to be more potent for the selected molecules, exhibiting five pharmacophoric features. The active scaffolds were further evaluated for enzyme inhibition efficacy against alleged receptor Pf‐DHFR computationally and in vitro, proving their candidature as lead dihydrofolate reductase inhibitors, and the selectivity of the test candidates was ascertained by toxicity study against Vero cells. Good oral bioavailability was also proved by studying pharmacokinetic properties.     

Dehydro-digitoxosides of digitoxigenin: Formation and importance for the digitoxin metabolism in the rat

Summary Only in the presence of NADPH and oxygen digitoxosides of digitoxigenin can be metabolized by rat liver microsomes. The metabolism includes 12-hydroxylation, formation of digitoxosides less the terminal digitoxose and of lipophilic metabolites. The lipophilic metabolites of digitoxin (dt-3), digitoxigeninbis-digitoxoside (dt-2), and of — mono-digitoxoside (dt-1) are formed by oxidation of the axial OH-group of the terminal digitoxosyl yielding the corresponding dehydro-digitoxosides. The structure could be confirmed by comparison with the synthetic compounds 15-dehydro-dt-3, 9-dehydro-dt-2, and 3-dehydro-dt-1, respectively.The terminal dehydro-digitoxosyl can be split off by liver microsomes even in the absence of NADPH. However, no further cleavage of the resulting digitoxosides could be detected. A cleavage rate of 2.8 nmoles/mg microsomal protein × 30 min was observed for both 15-dehydro-dt-3 and 9-dehydro-dt-2 (substrate concentration 50 M). In contrast to that, only 0.4 nmole 3-dehydro-dt-1 was cleaved under equal conditions. For the cleavage of 15-dehydro-dt-3 an apparent K _m of 200 M and a V _max of 440 pmoles dt-2 formed per mg microsomal protein x min was measured.Our results indicate that not the native but only the dehydro-digitoxosides can be cleaved. Therefore, two successive monoxygenase catalysed oxidations are necessary for the cleavage of the sugar chain of dt-3 before dt-1, the main substrate of conjugation enzymes, can be formed. Moreover, digitoxigenin will not be formed because of the high stability of 3-dehydro-dt-1.     

加替沙星合成方法的改进

以1-环丙基-6,7,8-三氟-1,4-(2H)-4-氧代喹啉-3-羧酸乙酯为原料,经过水解、缩合和甲氧基化制得加替沙星.此法成本低、步骤少、操作简便、条件温和,适于工业生产.     

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。     

N-(顺式-4-异丙基环己基-1-甲酰基)-D-苯丙氨酸和N-(反式-4-异丙基环己基-1-甲酰基)-L-苯丙氨酸的合成

目的合成N-(顺式-4-异丙基环己基-1-甲酰基)-D-苯丙氨酸和N-(反式-4-异丙基环己基-1-甲酰基)-L-苯丙氨酸.方法以(4-异丙基)环己基甲酸为原料,在二环己基碳二亚胺(DCC)作用下,与N-羟基琥珀酰亚胺反应得到(4-异丙基)环己基甲酸琥珀酰亚胺酯(3),柱色谱分离化合物3得到顺式和反式异构体.顺式体与D-苯丙氨酸甲酯发生酰化反应,碱水解后即得到N-(顺式-4-异丙基环己基-1-甲酰)-D-苯丙氨酸;而反式体与L-苯丙氨酸甲酯发生酰化反应,水解后得到N-(反式-4-异丙基环己基-1-甲酰)-L-苯丙氨酸.结果与讨论成功合成了目标化合物,反应总收率分别为39%和31%.     

1, 5-二芳基取代-1, 2, 4-三唑类衍生物的合成、 抗炎活性及其构效关系研究

摘 要：目的 寻找高活性的选择性环氧合酶-2抑制剂，探讨其构效关系。方法 根据已上市的选择性环氧合酶-2抑制剂celecoxib的构效关系以及分子模拟研究的结果，设计了两类1,5-二芳基取代-1,2,4-三唑类衍生物，以1-(4-取代苯基)-3-硫代氨基脲为原料，经多步反应合成目标化合物；用小鼠二甲苯致炎模型对目标化合物进行体外抗炎活性测试。结果与结论 合成了22个未见文献报道的新化合物，所有目标化合物的结构经IR、¹H-NMR、MS谱和元素分析确证。生物活性研究表明，化合物I₃、I₉、I₁₂、I₁₅、II₂、II₃、II₆、II₇具有较强的抗炎活性（P<0.01），其中化合物II₂、 II₃、 II₆的抗炎活性最强。构效关系分析发现，在三唑环1位芳基的对位引入F、Cl、Br吸电子基团以及3位引入磺酰基和甲硫基，对抗炎活性有重要的意义；在三唑环5位芳基的对位引入氨磺酰基，对抗炎活性有较大影响。     

具有抗癌活性的喜树碱类化合物的构效关系研究

目的 探寻喜树碱类似物的电子结构与抗癌活性的关系。方法 应用分子力学MM 法、量子化学从头算法和AMl法对18个喜树碱类似物进行几何优化，计算了化合物的电子结构，并应用逐步回归方法探寻化合物量子化学指数和抗癌活性的关系。结果 (1)喜树碱类似物的1og P与活性参数间呈抛物线关系；(2)分子的log P及20位碳原子电荷是影响化合物抗癌活性的重要因素；7位取代基的疏水参数1og P2对分子的log P影响很大；(3)得到较显著的QSAR方程：pIC50＝38．123 8．0907log P—0．81425(log P)^2—682．379QC20。结论 疏水性对喜树碱类似物的活性影响较大；D环及相连的羰基氧O23可能是与受体作用的活性中心；根据所得的QSAR及方程可以较准确地预测喜树碱类似物地活性。     

Differences in the uptake of cadmium and mercury by rat hepatocyte primary cultures. Role of a sulfhydryl carrier

Studies measuring the uptake of cadmium or mercury in isolated hepatocytes demonstrated that hepatocytes accumulated more cadmium than mercury in serum-containing medium, serum-free medium, or balanced salt solution. The preferential hepatocellular accumulation of cadmium, independent of medium composition, suggested that the uptake mechanism for cadmium and mercury might be different in hepatocytes. Pretreatment of hepatocytes with 50 microM N-ethylmaleimide decreased cadmium uptake by 23% while having no effect on the uptake of mercury was inhibited in a concentration-dependent manner. The uptake of cadmium was maximally inhibited (80%) with 75 microM parachloromercuribenzenesulfonate or 20 microM mercury respectively. Cadmium had no effect on mercury. Hepatocytes treated with parachloromercuribenzenesulfonate or mercury accumulated cadmium at a rate closely resembling the rate of mercury uptake in untreated hepatocytes. These results suggested that an SH-containing carrier may be operative in the uptake of cadmium by hepatocytes. Mercury can interact with this carrier to inhibit cadmium uptake; however, this carrier does not appear to facilitate mercury uptake.     

Lenvatinib for the treatment of renal cell carcinoma

Introduction: Renal cell carcinoma (RCC) accounts for 2–3% of all solid tumors. Expression of the receptor for the vascular endothelial growth factor (VEGF) is one of the most common features of RCC.

Areas covered: Lenvatinib is a novel multi-kinase inhibitor that has been studied in several solid tumors. It has shown promising results in the treatment of RCC, especially when combined with everolimus, In this review, we summarize the available data of lenvatinib for the treatment of advanced/metastatic renal cell carcinoma.

Expert opinion: Lenvatinib in combination with everolimus has provided encouraging results in both clinical and laboratory investigations showing that blocking angiogenesis and the mTOR signalling pathway could be a remarkable approach for treating RCC. As an additive to this type of approach it would be interesting in future clinical settings testing also the combination of lenvatinib and everolimus with immune-therapy.     

KISS-1及P16NK4A在宫颈鳞癌中的表达及意义

目的探讨KISS-1和P16NK4A在正常宫颈粘膜上皮、宫颈上皮内瘤变及宫颈鳞癌组织中的表达及其意义。方法采用免疫组化SP法检测40例正常宫颈粘膜上皮、40例子宫颈上皮内瘤样病变（cervical intraepithelial neoplasia,CIN）及40例宫颈鳞癌患者组织KISS—1及P16NK4A的表达。结果KISS-1和P16NK4A的阳性表达率在正常宫颈粘膜上皮、CIN及宫颈鳞癌中的表达率分别为85．0％、62．5％、29．7％和10．0％、65．0％、87．5％;KISS—1蛋白的失表达与宫颈鳞癌的浸润深度及淋巴结转移有关（P〈0．05）;P16NK4A的高表达与宫颈鳞癌的浸润深度及淋巴结转移有关（P〈0．05）;子宫颈鳞癌组织中KISS-1和P16NK4A的表达呈负相关（P〈0．01）。结论KISS-1的低表达和P16NK4A过表达与宫颈癌的发生、发展、浸润、转移有关。两者的表达负相关,联合检测KISS-1和P16NK4A表达可以在一定程度上预测宫颈癌患者的预后。     

Effects of aripiprazole and its active metabolite dehydroaripiprazole on the activities of drug efflux transporters expressed both in the intestine and at the blood-brain barrier

The inhibition potencies of aripiprazole and its active metabolite, dehydroaripiprazole, on the activities of human multidrug resistance protein 1 (MDR1/ABCB1; P‐glycoprotein), breast cancer resistance protein (BCRP/ABCG2) and multidrug resistance‐associated protein 4 (MRP4/ABCC4), that are drug efflux transporters expressed both in the intestine and at the blood–brain barrier (BBB), were investigated. Aripiprazole and dehydroapripiprazole showed relatively strong inhibitory effects on human MDR1 with IC₅₀ values of 1.2 and 1.3 µm in human MDR1‐transfected Mardin‐Darby canine kidney (MDCKII‐MDR1) cells, respectively. The inhibition potencies of other atypical antipsychotics (risperidone, paliperidone, olanzapine and ziprasidone) for human MDR1 were also evaluated using the same in vitro experimental system and IC₅₀ values were more than 10‐fold higher than those of the two compounds. Aripiprazole and dehydroaripiprazole also had inhibition potencies against human BCRP with IC₅₀ values of 3.5 and 0.52 µm , respectively. The ratios of steady‐state unbound concentrations of aripiprazole and dehydroaripiprazole to their IC₅₀ values against human MDR1 and BCRP activities were less than 0.1, whereas the theoretically maximum gastrointestinal concentration of aripiprazole ([I]₂) to its IC₅₀ values was much higher than the cut‐off value of 10, proposed by the International Transporter Consortium (ITC) and the Food and Drug Administration (FDA). In contrast, aripiprazole and dehydroaripiprazole showed almost no inhibitory effect against the activity of human MRP4. These findings indicate that aripiprazole is unlikely to cause drug–drug interactions (DDIs) at the BBB when co‐administered with substrate drugs of these drug transporters investigated. However, interactions at the intestinal absorption process may be of concern. Copyright © 2012 John Wiley & Sons, Ltd.     

基于GEO和TCGA数据库筛选恶性胸膜间皮瘤免疫治疗疗效预测关键基因

目的 探索免疫治疗在恶性胸膜间皮瘤（malignant pleural mesothelioma,MPM）中免疫疗效相关关键分子及其可能的机制。方法 2018年7月至2020年7月,纳入GEO数据库中GSE117358的表达谱进行分析;首先,根据是否对免疫治疗是否有效分为两组：免疫治疗反应组和免疫治疗无反应组,同时分析两组之间差异有统计学意义基因;其次,分析两组差异有统计学意义基因在基因本体（gene ontology,GO）生物学行为及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes,KEGG）富集信号通路等方面的差异;最后,对差异有统计学意义基因构建蛋白-蛋白互作网络,筛选在生物学行为中的重要模块及关键基因,并对关键基因在TCGA数据库中进行整合分析验证。结果 在免疫治疗反应组（12个样本）及免疫治疗无反应组（12个样本）中共筛选出1 025个差异基因,相对于免疫治疗无反应组,在免疫治疗反应组中有782个上调和243个下调基因。GO和KEGG分析显示,这些差异基因的功能主要集中在细胞因子受体通路、细胞黏附通路、T细胞受体通路及...     

Molecular modulation of the copper and cisplatin transport function of CTR1 and its interaction with IRS-4

The copper influx transporter CTR1 is also a major influx transporter for cisplatin (cDDP) in tumor cells. It influences the cytotoxicity of cDDP both in vivo and in vitro. Whereas Cu triggers internalization of CTR1 from the plasma membrane, cDDP does not. To investigate the mechanisms of these effects, myc-tagged forms of wild type hCTR1 and variants in which Y103 was converted to alanine, C189 was converted to serine, or the K178/K179 dilysine motif was converted to alanines were re-expressed in mouse embryo cells in which both alleles of CTR1 had been knocked out and also in HEK293T cells. The Y103A mutation and to a lesser extent the C189S mutation reduced internalization of CTR1 induced by Cu while the K178A/K179A had little effect. Both Y103 and C189 were required for Cu and cDDP transport whereas the K178/K179 motif was not. While Y103 lies in an YXXM motif that, when phosphorylated, is a potential docking site for phosphatidylinositol 3-kinase and other proteins involved in endocytosis, Western blot analysis of immunoprecipitated myc-CTR1, and proteomic analysis of peptides derived from CTR1, failed to identify any basal or Cu-induced phosphorylation. However, proteomic analysis did identify an interaction of CTR1 with IRS-4 and this was confirmed by co-immunoprecipitation from HEK cells expressing either FLAG-CTR1 or myc-CTR1. The interaction was greater in the Y103A-expressing cells. We conclude that Y103 is required for the internalization of hCTR1 in response to Cu, that this occurs by a mechanism other than phosphorylation and that mutation of Y103 modulates the interaction with IRS-4.