新型苯并吗啉-8-甲酰胺衍生物的合成及抗肿瘤活性

以6-溴N-(4,6-二甲基-2-氧代-1,2-二氢吡啶-3-甲基)苯并吗啉-8-甲酰胺为原料,经N-磺酰化和Suzuki偶联反应两步合成17个新型的6-芳基取代N-磺酰基苯并吗啉-8-甲酰胺衍生物(3a~3q),其结构经¹H NMR、¹³C NMR和MS(EI)表征。采用Alpha LISA法对化合物进行EZH2(WT)酶活性检测;采用MTT法测试了化合物对非小细胞肺癌细胞A549和人类B细胞淋巴瘤细胞SU-DHL-4的细胞增殖抑制作用。结果表明：化合物3j的浓度为1μmol·L^-1时,对EZH2抑制率为68%; 对A549细胞和SU-DHL-4细胞的IC₅₀值分别达到10.4 μmol·L^-1和2.8 μmol·L^-1。      

N-2-氯苄基苯甲酰胺的固相合成及其晶体结构分析

N-2-氯苄基苯甲酰胺采用固相法制备.聚苯乙烯磺酰氯(1)在吡啶存在下与2-氯苄胺反应得到N-2-氯苄基磺酰胺树脂(2),进一步在吡啶催化下用苯甲酰氯进行酰化得到N-2-氯苄基-N-苯甲酰基磺酰胺树脂(3).用TiCl₄/Zn/THF试剂处理,从树脂3上解脱得到产物N-2-氯苄基苯甲酰胺(4),产率92%.N-2-氯苄基苯甲酰胺的晶体属于单斜晶系,P2₁空间群,晶胞参数:a=0.7149(3)_nm,b=0.8772(4)_nm,c=0.9704(4)_nm;β=95.172(7)°;晶体结构中存在两种分子间氢键相互作用.     

N-(2-对甲苯磺酰胺基乙基)单氮杂冠醚的合成及其晶体结构

N-对甲苯磺酰基乙二胺分别与1,11-二碘-3,6,9-三氧杂十一烷和1,14-二碘-3,6,9,12-四氧杂十四烷反应,制得N-(2-对甲苯磺酰胺基乙基)单氮杂-12-冠-4或15-冠-5;当用N-(2-对甲苯磺酰胺基乙基)二乙醇胺与1,11-二对甲苯磺酸酯-3,6,9-三氧杂十一烷反应时,才能获得N-(2-对甲苯磺酰胺基乙基)单氮杂-18-冠-6.从甲醇溶液中培养得N-(2-对甲苯磺酰胺基乙基)单氮杂-12-冠-4单晶,属单斜晶系,P2₁/a空间群;a=1.4229(1)nm,b=0.9595(2)nm,c=1.4564(1)nm,β=102.20(1)°,V=1.9435nm³,Z=4.最终偏离因子R=0.043.     

N-(4-取代嘧啶-2-基)苄基磺酰脲和苯氧基磺酰脲的3D-QSAR研究

采用比较分子力场分析(CoMFA)方法,对两类单取代嘧啶类似物、6个N-(4-取代嘧啶-2-基)-2-甲氧羰基苄基磺酰脲(1a～1f)和14个N-(4-取代嘧啶-2-基)-2-取代苯氧基磺酰脲(2a～2n)进行三维定量构效关系(3D-QSAR)研究.建立了一个较为可靠的预测模型.结果表明,分子中苯环邻位、嘧啶环形成氢键的N原子处以及嘧啶环4位和6位附近负电荷增加;苯环邻位乙氧基的CH₂CH₃附近选择带正电的原子;苯环邻位乙氧基附近空间体积增加,而嘧啶环4位甲氧基稍远处取代基的立体位阻不超过此位置,将有利于提高活性.最后解释了修饰磺酰脲的除草剂仍具有较高活性的原因.     

新型二氢卟吩-5-氟尿嘧啶的合成

以系列羟基卟啉为原料,用对甲苯磺酰肼还原,再用四氯对苯醌氧化,制得二氢卟吩(2a~2d);以二环己基碳二亚胺为脱水剂,2与5-氟尿嘧啶-1-基乙酸经酯化反应合成了4个新型二氢卟吩-5-氟尿嘧啶化合物(3a~3d),总收率分别为6.9%, 6.9%, 7.0%和7.6%,其结构经UV-Vis, ¹H NMR, IR和MS表征。     

三氟甲基取代叔丁基亚磺酰亚胺的不对称aza-Morita-Baylis-Hillman反应

三氟甲基取代的N-叔丁基亚磺酰亚胺和α, β-不饱和羰基化合物在三乙烯二胺(DABCO)和Ti(OⁱPr)₄共催化下反应，实现了底物诱导的不对称aza- Morita-Baylis-Hillman反应，以高产率(49%~97 %)，高非对映选择性(dr>99/1)合成了一系列N-叔丁基亚磺酰基-α-甲亚基-β-三氟甲基-β-氨基酸酯产物。该产物经脱保护、关环可进一步转化为光学纯的α-甲亚基-β-三氟甲基-β-内酰胺类化合物，产物结构经¹H NMR、 ¹³C NMR和HR-MS确证。      

三氟甲基嘧啶磺酰脲的合成与除草活性

以三氟乙酰乙酸乙酯和盐酸胍为原料,经环合、氯化、甲氧基化、胺化反应合成了4种2-氨基-4-取代-6-三氟甲基嘧啶中间体(Ⅰa~Ⅰd),Ⅰ与邻甲氧羰基苯磺酰异氰酸酯亲核加成,合成了3种含不同取代嘧啶环的磺酰脲化合物(Ⅱb~Ⅱd)。其中2-氨基-4-二甲胺基-6-三氟甲基嘧啶(Ⅰd)和N[-2-(′4-二甲胺基-6-三氟甲基)嘧啶基]-2-甲氧羰基苯磺酰脲(Ⅱd)为新化合物,结构经1H NMR和MS表征。初步除草活性测定结果表明,N-[2-′(4-甲氧基-6-三氟甲基)嘧啶基]-2-甲氧羰基苯磺酰脲(Ⅱc)在浓度为200 ga.i./ha时对稗草和马唐的防效分别为70%和50%。     

N-二异丙基磷酰基-N-[2-(3',4'-亚甲二氧苯基)]乙基甘氨酸的单昌结构的研究

三类杉酯碱是一种具有显著抗肿瘤活性的化合物, 在以磷脱基, 磺酰基乙酰基作N-苯乙基甘氨酸上氨基的保护基进行分子内酰化的Friede-Crafts反应, 合成三尖杉酯碱母核-(N-7,8-亚甲二氧苯并-3-氮杂环庚酮-1)时所得结果相差甚远, 特别是用磷酰基保护甘氨酸衍生物可以避免脱羰, 顺利地进行内酰化反应, 对合成三尖杉酯碱的母核有重要的意义。为了阐明磷酰基对氨基酸中氮原子上的弧对电子的稳定作用, 我们测定了N-二异丙基磷酰基-N-[2-(3',4'-亚甲二氧苯基)]乙基甘氨酸的单晶结构。     

N-二异丙基磷酰基-N-[2-(3',4'-亚甲二氧苯基)]乙基甘氨酸的单昌结构的研究

三类杉酯碱是一种具有显著抗肿瘤活性的化合物, 在以磷脱基, 磺酰基乙酰基作N-苯乙基甘氨酸上氨基的保护基进行分子内酰化的Friede-Crafts反应, 合成三尖杉酯碱母核-(N-7,8-亚甲二氧苯并-3-氮杂环庚酮-1)时所得结果相差甚远, 特别是用磷酰基保护甘氨酸衍生物可以避免脱羰, 顺利地进行内酰化反应, 对合成三尖杉酯碱的母核有重要的意义。为了阐明磷酰基对氨基酸中氮原子上的弧对电子的稳定作用, 我们测定了N-二异丙基磷酰基-N-[2-(3',4'-亚甲二氧苯基)]乙基甘氨酸的单晶结构。     

白杨素甘氨酸类化合物的合成及抗癌活性

以白杨素为起始原料, 通过卤代和水解反应制得中间产物7-O-羧烷基化的白杨素衍生物(6~9); 然后以1-乙基-3-(3-二甲氨基丙基)碳二亚胺(EDCI)、 1-羟基苯并三氮唑(HOBt)和4-二甲氨基吡啶(DMAP)为催化体系, 4个中间产物分别与甘氨酸甲酯盐酸盐进行酰胺缩合反应, 制得白杨素甘氨酸甲酯类化合物12~15; 化合物12~15在pH=10~11和室温下水解得到相应的白杨素甘氨酸类化合物(16~19). 所有目标化合物的结构均经 ¹H NMR, ¹³C NMR, IR以及MS确认. 以顺铂为阳性对照药物, 采用噻唑蓝比色(MTT)法检测了目标化合物对人肝癌细胞HepG2和人胃癌细胞MGC-803的体外增殖抑制作用. 结果表明, 目标化合物14~16, 18和19的体外抗肿瘤活性明显强于白杨素, 且化合物18(IC₅₀=4.36 μmol/L)对MGC-803细胞的增殖抑制作用强于阳性药物顺铂(IC₅₀=4.40 μmol/L).     

高效液相色谱法同时测定血清和尿中厚朴酚与和厚朴酚

 建立了大鼠服用厚朴提取物后的血清中及尿中厚朴酚与和厚朴酚的高效液相色谱测定法。色谱柱填料为SpherisorbC18,流动相为甲醇-水-冰醋酸(体积比为70∶30∶1),UV检测波长为294nm,灵敏度0.005AUFS。样品用甲醇沉淀蛋白,上清液酸化后用乙酸乙酯-乙醚萃取,然后测定其中的药物浓度。血清和尿中的药物浓度与峰面积的线性关系良好,线性范围分别为0.05～2mg/L(厚朴酚)、0.025～1mg/L(和厚朴酚);精密度和重现性良好。血清中厚朴酚与和厚朴酚的平均加样回收率分别为95.6%(RSD=3.85%)和93.8%(RSD=3.95%),尿中分别为96.0%(RSD=3.83%)和94.9%(RSD=3.54%)。     

Voltammetric detection of magnolol in Chinese medicine based on the enhancement effect of mesoporous Al/SiO(2)-modified electrode

The enhancement effect of mesoporous Al-doped silica (Al/SiO(2))-modified electrode was investigated. Due to the properties such as large surface area, strong adsorptive ability and numerous active sites, mesoporous Al/SiO(2) alters the structure and property of electrode/solution interface, then greatly improves the electrochemical response of magnolol. The electrochemical behavior of magnolol was examined in detail. It is found that the oxidation peak current of magnolol remarkably increases at the mesoporous Al/SiO(2)-modified electrode. Based on this, a sensitive and convenient electrochemical method was developed for the determination of magnolol. The linear range is over the range from 7.5 x 10(-8) to 2.0 x 10(-5) mol L(-1). The limit of detection (S/N=3) is as low as 2.5 x 10(-8) mol L(-1). Finally, this novel method was successfully used to determine the magnolol in Chinese traditional medicines.     

Toxicity study of allelochemical-like pesticides by a combination of 3D-QSAR, docking, Local Binding Energy (LBE) and GRID approaches

3D-QSAR, Docking, Local Binding Energy (LBE) and GRID methods were integrated as a tool for predicting toxicity and studying mechanisms of action. The method was tested on a set of 73 allelochemical-like pesticides, for which acute toxicity (LD(50)) for the rat was available. 3D-QSAR gave a model with high predictive ability and the regression maps indicated the important toxic chemical substituents. Significant ligand-protein residue interactions and oxidation positions in the binding site were found by docking analysis using CYP1A2 homology modelling. The binding energies of the compounds and the important substituents (Local Binding Energy, LBE) were calculated in order to demonstrate quantitatively the substituent contributions in the metabolism and toxicity. The GRID examination identified the CYP1A2 binding pocket feature. Finally, a 3D-QSAR map was compared to the GRID map, showing good overlaps and confirming the important role of CYP1A2 in allelochemical-like compounds toxicity.     

Analysis of magnolol and honokiol in biological fluids by capillary zone electrophoresis

Capillary zone electrophoresis (CZE) method was used for analysis of magnolol and honokiol. Under the optimized condition, CZE with UV absorption detection provided that the limit of detection was at microM level. To enhance detection sensitivity of magnolol and honokiol, CZE separation system was coupled with a laser-induced fluorescence (LIF) detector for the first time. The limits of detection of magnolol and honokiol were 12 nM (3.20 ng ml(-1)) and 18 nM (4.79 ng ml(-1)), respectively, showing that the CZE-LIF system provides greater than 100-fold sensitivity improvements than does the CZE-UV system. The developed method was applied to analyze magnolol and honokiol in spiked human plasma samples, microsome incubation samples as a preliminary demonstration of its potential in pharmacokinetic studies.     

2D SMARTCyp reactivity-based site of metabolism prediction for major drug-metabolizing cytochrome P450 enzymes

Cytochrome P450 (CYP) 3A4, 2D6, 2C9, 2C19, and 1A2 are the most important drug-metabolizing enzymes in the human liver. Knowledge of which parts of a drug molecule are subject to metabolic reactions catalyzed by these enzymes is crucial for rational drug design to mitigate ADME/toxicity issues. SMARTCyp, a recently developed 2D ligand structure-based method, is able to predict site-specific metabolic reactivity of CYP3A4 and CYP2D6 substrates with an accuracy that rivals the best and more computationally demanding 3D structure-based methods. In this article, the SMARTCyp approach was extended to predict the metabolic hotspots for CYP2C9, CYP2C19, and CYP1A2 substrates. This was accomplished by taking into account the impact of a key substrate-receptor recognition feature of each enzyme as a correction term to the SMARTCyp reactivity. The corrected reactivity was then used to rank order the likely sites of CYP-mediated metabolic reactions. For 60 CYP1A2 substrates, the observed major sites of CYP1A2 catalyzed metabolic reactions were among the top-ranked 1, 2, and 3 positions in 67%, 80%, and 83% of the cases, respectively. The results were similar to those obtained by MetaSite and the reactivity + docking approach. For 70 CYP2C9 substrates, the observed sites of CYP2C9 metabolism were among the top-ranked 1, 2, and 3 positions in 66%, 86%, and 87% of the cases, respectively. These results were better than the corresponding results of StarDrop version 5.0, which were 61%, 73%, and 77%, respectively. For 36 compounds metabolized by CYP2C19, the observed sites of metabolism were found to be among the top-ranked 1, 2, and 3 sites in 78%, 89%, and 94% of the cases, respectively. The computational procedure was implemented as an extension to the program SMARTCyp 2.0. With the extension, the program can now predict the site of metabolism for all five major drug-metabolizing enzymes with an accuracy similar to or better than that achieved by the best 3D structure-based methods. Both the Java source code and the binary executable of the program are freely available to interested users.     

Liquid chromatographic determination of honokiol and magnolol in hou po (Magnolia officinalis) as the raw herb and dried aqueous extract

A validated analytical method is described for the determination of honokiol and magnolol in Hou Po (Magnolia officinalis) as the dried raw herb and the commercially prepared dried aqueous extract. The samples were extracted with methanol by the Soxhlet method, and the extract was analyzed by liquid chromatography with photodiode array (LC/PDA) detection with confirmation of analyte identity by negative-ion electrospray ionization tandem mass spectrometry (ESI-MS/MS). A C18 column was used with a menthanol--0.1% aqueous acetic acid gradient mobile phase. Honokiol and magnolol were quantified at 288 nm. With the MS detector, the honokiol precursor ion at m/z 265 was shown to produce ions at m/z 222 and 224. For magnolol, the precursor ion at m/z 265 produced the ions at m/z 247 and 245. Comparable results were obtained for the LC/PDA and LC/ESI-MS/MS methods of quantitation. Six commercially prepared dried aqueous extracts were analyzed. The levels of honokiol and magnolol found in the raw herb were 17.0 and 21.3 mg/g, respectively. The limits of detection for honokiol and magnolol in the raw herb were 0.45 and 0.58 mg/g, respectively, and in the dried aqueous extract, 0.04 and 0.30 mg/g, respectively.     

High-performance liquid chromatographic method for simultaneous determination of honokiol and magnolol in rat plasma

A high-performance liquid chromatographic (HPLC) method is described for the simultaneous determination of honokiol and magnolol in rat plasma. The plasma was deproteinized with acetonitrile which contained an internal standard (diphenyl) and was separated from the aqueous layer by adding sodium chloride. Honokiol and magnolol are extracted into the acetonitrile layer with high yield, and determined by reversed-phase HPLC and ultraviolet detection. The limits of quantitation for honokiol and magnolol were 13 and 25 ng ml⁻¹ in plasma, respectively, and recovery of both analytes was greater than 93%. The assay was linear from 20 to 200 ng ml⁻¹ for honokiol and from 40 to 400 ng ml⁻¹ for magnolol. Variation over the range of the standard curve was less than 15%. The method was used to determine the concentration-time profiles of honokiol and magnolol in the plasma following rectal administration of Houpo extract at a dose of 245 mg kg⁻¹, equivalent to 13.5, 24.4 mg kg⁻¹ of honokiol and magnolol, respectively.     

Determination of Magnolol and Honokiol by Non-aqueous Capillary Electrophoresis

Introduction Capillaryelectrophoresis(CE),ahighefficiency separationtechnique,hasrapidlydevelopedsince1981[1].Notonlyhavedifferentmodesbeenrepor ted[1—3],butalsosomenon aqueouselectrophoresis media[4]havebeenapplied.Theimportantproperties ofnonaqueousmed…     

Determination of honokiol and magnolol by micro HPLC with electrochemical detection and its application to the distribution analysis in branches and leaves of Magnolia obovata

A simple and sensitive method has been developed for determining honokiol and magnolol in fresh Magnolia obovata (M. obovata) by micro high-performance liquid chromatography with electrochemical detection (microHPLC-ECD). Chromatography was performed using a Capcell Pak C-18 UG 120 microbore octadecylsilica (ODS) column, methanol-water-phosphoric acid (65 : 35 : 0.5, v/v/v), as a mobile phase and applied potential at +0.8 V vs. Ag/AgCl. Peak heights were found linearly related to the amounts of honokiol and magnolol injected from 0.67 pg to 2.0 ng (r>0.999). The detection limits (S/N=3) were 0.13 pg, respectively. Honokiol and magnolol of 0.27 ng were detected with relative standard deviation (RSD) of 0.73 and 1.17% (n=5), respectively. Honokiol and magnolol in Magnolia Bark of the Japanese Pharmacopoeia were extracted with 70% methanol, diluted with a mobile phase, and injected into the microHPLC-ECD for determination. Recoveries of honokiol and magnolol in Magnolia Bark exceeded 98.7% with RSD, less than 0.93% (n=5). Determination of the distributions of honokiol and magnolol in bark, phloem, wood, leaf blades, and petioles of fresh M. obovata were made using weight samples of 40-238 mg. This method is useful to determine honokiol and magnolol in M. obovata, which is a candidate for crude magnolia bark for traditional Japanese herbal medicines.     

The inhibition of eugenol on glucan is essential for the biofilm eradication effect on caries-related biofilm in an artificial mouth model

This study aimed to evaluate the inhibitory effects of herbal extracts on caries-related bacteria, glucan and biofilm in?vitro. Sensitive tests of bacteria were carried out by broth dilution method on a 96-microwell plate. Glucan inhibition tests were carried out using the phenol sulphate method. A minimum biofilm inhibitory concentration (MBIC) test was performed in an artificial mouth model. The results of the MBIC of agents were 8, 16 and 32?mg?mL(-1) for eugenol, tannic acid and magnolol, respectively. For the results of glucan inhibition tests, over 63%, 28% and 27% inhibition occurred on insoluble glucan syntheses of Streptococcus sobrinus for eugenol, magnolol and tannic acid, respectively. Over 46%, 16% and 13% inhibition on soluble glucan syntheses for eugenol, magnolol and tannic acid, respectively, were also observed. In conclusion, the inhibition of eugenol on glucan is essential for the biofilm inhibition effect on caries-related biofilm in an artificial mouth model in?vitro.