三种非甾体类抗炎药与脂质体的相互作用

以卵磷脂脂质体为生物膜模型, 分别采用二阶导数吸收光谱法和以1-苯胺基-8-萘磺酸铵(ANS)为探针的荧光光谱法, 研究了三种丙二酸类非甾体抗炎药吲哚美辛、舒林酸和托美丁与脂质体的相互作用. 药物的二阶导数吸收谱成沟槽型,在脂质体中沟槽变浅, 但波长基本上不移动, 表明药物结合在磷脂双层的表面而没有进入到脂双层内部. 在荧光光谱中, 抗炎药可以结合到脂质体上, 游离出ANS, 猝灭ANS 在脂质体中的强烈荧光, 表观猝灭常数K_SV^app＞10^12 L·mol^(-1); 药物先行结合到脂质体上后, 会降低ANS结合到脂质体上的机率. 同样表明药物可以结合到脂质体表面磷脂分子的头部, 结合能力的强弱与吸收谱得到的结果一致, 舒林酸最强, 吲哚美辛其次, 托美丁的结合能力较弱.     

光谱法研究吲哚美辛,舒林酸及其金属配合物与牛血清蛋白的相互作用

以曙红Y(Eosin Y)为探针,采用竞争结合法考查了非甾体抗炎药吲哚美辛和舒林酸及其相关的金属配合物与BSA的结合。在中性环境中,曙红Y的吸收光谱随着BSA的加入发生明显变化,结果表明,曙红Y与BSA之间的结合以静电作用为主,两者之间可能发生电子转移。曙红Y与BSA结合后,抗炎药吲哚美辛,舒林酸的加入可以破坏这种结合,曙红Y部分游离出来,其吸收值恢复,说明药物可以同样的方式结合到BSA上,抢占曙红Y在BSA上的结合位点。将药物转化为金属配合物后,与血清蛋白的亲和程度更强,铜(Ⅱ)的配合物最为突出,推测将非甾体抗炎药转化为铜配合物后,有更好的药理活性。     

一种不对称菁染料及其与不同结构DNA的相互作用

为考察小分子配基与不同核酸结构的结合机理,发展新的核酸探针分子,合成了一种新型一次甲基不对称菁染料(MTP).吸收光谱、荧光光谱及圆二色光谱研究结果表明,MTP可与平行和混合平行G-四链体DNA(如c-myc和22AGK+)较强地结合,并引起130~180倍的荧光增强;其与单/双链DNA作用较弱,导致40~60倍荧光增强;而与反平行G-四链体DNA(如TBA和22AGNa+)的作用最弱,只引起15~25倍荧光增强;以上结果表明MTP可作为荧光探针分子用于区别不同结构的核酸分子.结合机理研究表明,平行和混合平行G-四链体DNA优先通过沟槽结合模式结合1分子MTP,再通过末端堆积模式结合另1分子MTP.     

中性介质中中性红与双链DNA作用的光谱

通过分子吸收、荧光发射和共振光散射测定,表征了在水溶液介质中中性红(NR)与双 螺旋DNA的作用.在pH 7.63和离子强度低于0.01的水溶液介质中,随着NR与DNA的摩尔比(R)变 化,存在有两种结合方式.第一种结合方式发生在R > 2.22,此时获得共振光散射光谱增强信 号,表明NR在DNA分子表面发生聚集,集聚特性可使用RLS测定数据进行Scatchard分析;第二种 结合方式发生在R < 2.22,此时NR内嵌到DNA分子的双链碱基对之间,具有特征波长红移和分 子吸收增色效应,发生了从DNA到NR的分子能量转移,能观察到荧光增强.     

水溶液微悬浮聚合法制备酸性药物吲哚美辛分子印迹微球及其色谱表征

以带有羧基的酸性药物吲哚美辛为模板分子、碱性的4-乙烯基吡啶为功能单体,采用水溶液微悬浮聚合法制备了用于色谱分离的微米级分子印迹微球.详细讨论了流动相中缓冲溶液的pH值对吲哚美辛在MIMs柱上的容量因子(k′)、分离因子(α)和印迹因子(β)的影响.通过MIMs柱对吲哚美辛和4-氨基吡啶(4-AP)的保留行为的比较,证明以4-乙烯基吡啶为功能单体制得的MIMs对吲哚美辛的识别作用,主要靠吡啶环上氮原子与吲哚美辛羧基之间的离子键相互作用,以及吡啶环与模板分子之间的π-π相互作用.     

光谱法研究对氨基苯乙酮缩对二甲氨基苯甲酰腙与DNA的相互作用

合成了对氨基苯乙酮缩对二甲氨基苯甲酰腙(AAPABH),采用荧光光谱法和UV-Vis吸收光谱法研究了AAPABH与DNA的相互作用。在pH 7.4Tris-HCl缓冲溶液中,以345 nm激发该化合物发射弱荧光,加入DNA后出现明显的荧光增强现象,表明探针分子与DNA结合形成了稳定配合物,其结合常数为5.48×107L/mol。研究了离子强度对体系荧光强度的影响,结果显示NaCl的加入未引起AAPABH-DNA体系荧光强度明显变化,表明AAPABH和DNA间不存在静电作用;同时DNA对AAPABH吸收光谱的影响表现为减色效应、探针分子与热变性后的DNA作用力明显小于与未变性DNA间的作用力、探针分子与溴化乙锭(EB)竞争结合DNA使得EB-DNA体系荧光猝灭,上述实验结果均表明探针分子主要以嵌插作用方式与DNA作用。     

甲基苯丙胺与血清白蛋白相互作用的光谱表征

采用荧光光谱和分子对接模拟研究了甲基苯丙胺与牛血清白蛋白(BSA)之间的相互作用,发现甲基苯丙胺对BSA的荧光有明显的猝灭作用.采用分子对接的方法模拟了甲基苯丙胺与BSA的分子动力学过程.结果显示,甲基苯丙胺可能与BSA中具有内源荧光特性的色氨酸和苯丙氨酸通过静电引力发生相互作用.利用荧光光谱进一步研究了甲基苯丙胺与氨基酸的相互作用.发现甲基苯丙胺对两种氨基酸的荧光都产生了明显的猝灭作用.研究结果表明,由于静电引力的作用,甲基苯丙胺与BSA发生了结合,结合位点是BSA中的色氨酸和苯丙氨酸.     

DNA与苯胺红T的相互作用与荧光定量检测

荧光染料作为分子探针用于核酸的定量测定和构象分析一直备受关注 [1] .已有数十种荧光染料被应用[2 ,3 ] ,如溴化乙锭早期常用作 DNA探针[4 ,5] ,因其有很强的致癌性 ,目前已很少采用 ,而那些无毒性或含有可被衍生的活性基团的探针分子越来越受到青睐 [6～ 8] .苯胺红 T(ST)是一种阳离子型荧光染料 ,它既可很容易地与 DNA双螺旋结构发生插入作用 ,又可由带正电的吩嗪环与 DNA上带负电的磷酸基发生强烈的静电吸附作用 ,还可利用其氨基与 DNA或其它生物分子进行交联 .He等 [9] 曾研究了它与小牛胸腺 DNA之间的作用 ,得到的结合位点数…     

米托蒽醌与B-DNA相互作用的分子模拟

针对嵌插型抗癌药物米托蒽醌(mitoxantrone,MTX)同B-DNA间作用模式的争议,采用分子模拟方法研究了米托蒽醌分子与B-DNA分子的相互作用.结果表明:米托蒽醌分子插入到B-DNA中有大小沟选择性及碱基对特异性,更倾向从小沟方向插入到DNA分子中;对5'-CG碱基对有特异性识别.通过详细能量项的分析,揭示了米托蒽醌插入DNA分子的驱动力及对碱基的特异性识别作用主要是空间相互作用特别是静电相互作用.在最佳作用位点复合物的构象分析则表明蒽醌环只有一部分插入碱基对中,侧链在小沟中延磷酸基骨架以3'-5'方向伸展,并通过静电作用进一步增强米托蒽醌与B-DNA的结合.     

双链DNA分子内电荷转移超交换机理

设计并合成了一系列寡聚核苷酸组成的双链DNA分子,通过检测样品中二氨基嘌呤(Ap)荧光峰强度和相对荧光量子产率来研究DNA分子内电荷转移.实验中直接分辨和观测到双链DNA分子内电荷转移超交换机理,超交换机理在近距离起作用；而电荷转移跳跃机理,可能是通过极子运动形式体现.     

Electron Transfer Processes in the Reactivity of Nonsteroidal Anti-inflammatory Drugs in the Ground and Excited States

The nonsteroidal anti-inflammatory drugs (NSAID), naproxen, sulindac and indomethacin, were shown to donate electrons to nitro blue tetrazolium (NBT) when irradiated with UV light in deoxygenated aqueous buffer solution (pH 7.4, 30°C). The reaction was monitored spec-trophotometrically by the appearance of the diformazan reduction product from NBT. The electron transfer process facilitates the decomposition of the drugs. Naproxen in the presence of NBT is photodegraded principally to the alcohol (2-[1-hydroxyethyl]-6-methoxynaphthalene) at a rate approximately 20-fold faster than when irradiated alone in deoxygenated conditions. The photoproduct from naproxen also participates in the electron transfer to NBT but at a much slower rate than naproxen. Irradiation of sulindac or indomethacin in the presence of NBT caused the slow photoreduction of NBT to diformazan. In the absence of NBT, indomethacin and sulindac are essentially unreactive when irradiated in aqueous solution. The ability of a number of NSAID to act as electron donors in their ground state was studied by observing their oxidation by potassium peroxodisulfate in pH 7.0 phosphate buffer at 50°C. The HPLC analysis of the drug remaining showed that the 2-arylpropionic acid NSAID (naproxen, ibuprofen, ketoprofen and suprofen) reacted at a rate equivalent to the thermal decomposition of peroxodisulfate. The major products were the same as detected in the photooxidation of these drugs, resulting from decarboxylation and oxygen addition but also included a dimeric compound. On the other hand, the NSAID that do not contain the propionic acid substituent all reacted more slowly with peroxodisulfate, enabling specific reaction rate constants to be evaluated.     

A temperature-dependent interaction of neutral red with calf thymus DNA

Neutral red (NR) is used as a probe to study the temperature and concentration dependent interaction of a cationic dye with nucleic acid. A temperature-dependent interaction of NR with calf thymus DNA (CT DNA) has been studied by differential pulse voltammetry (DPV), UV-Visible absorption, circular dichroism (CD) and fluorescence spectroscopy. The experimental results of increasing peak current, changes in the UV-Visible absorption and fluorescence spectra of NR and decreasing the induced circular dichroism (ICD) intensity show that (i) the binding mode of NR molecules is changed from intercalating into DNA base pairs to aggregating along the DNA double helix and (ii) the orientation of NR chromophore in DNA double helix is also changed with the temperature.     

Fluorescence quenching of ethidium ion by porphyrin cations and quaternary ammonium surfactants in the presence of DNA

Fluorescence quenching of free and DNA-bound ethidium bromide (EB) by a number of quaternary ammonium and other compounds was studied. For free EB or bound EB at lower DNA concentration the fluorescence quenching follows the Stern–Volmer equation and at higher DNA concentration follows an exponential model. At least at low quencher concentrations the quenching efficiency varies with DNA or NaCl concentrations and is about 100 times greater for bound than free EB. The quenching pathways may involve energy transfer and conformational loosening or distortion of the DNA helix in addition to possible electron transfer.     

Fluorescence studies on the interaction of ethidium bromide with duplex, triplex and quadruplex DNA structures

Under different conditions, oligonucleotides can form several alternative DNA structures such as duplex, triplex and quadruplex. All these structures can interact with ethidium bromide (EB) and make its fluorescence intensity change. The fluorescence spectra and other related parameters provided by static fluorescence techniques showed that the interaction mechanisms between EB and these structures were not always the same. Among them, B type duplex and triplex DNA adopt an intercalative mode when binding to the EB, which has a relatively high efficiency of energy transfer and the fluorescence of EB cannot be quenched easily. While for the parallel duplex DNA, the interaction mode is an outside binding in which energy transfer can hardly happen and its fluorescence intensity as well as Stern-Volmer constant is almost the same to the free EB. For the quadruplex, the binding mechanism to EB is more complex. Results from the energy transfer and quenching studies indicate that the two interaction modes note     

三聚氰胺对DNA潜在损伤作用的研究

在生理酸度条件下(pH 7.4),采用溴化乙锭(EB)为荧光探针的荧光光谱法、I-离子荧光猝灭效应、DNA熔点和粘度效应等手段,研究了三聚氰胺与DNA的相互作用。随着DNA的加入,三聚氰胺的荧光强度明显减小而且三聚氰胺能够猝灭DNA-EB复合物的荧光,说明三聚氰胺能够竞争置换EB而与DNA作用;三聚氰胺的加入使得DNA的粘度增大,DNA-EB的熔点降低;DNA的加入减小了I-对三聚氰胺荧光的猝灭程度。三聚氰胺以嵌插方式作用于DNA的亲核位点,意味着三聚氰胺进入生物体后有可能通过形成DNA加合物的形式造成DNA损伤,从而最终导致基因突变。     

Studies of interaction of emodin and DNA in the presence of ethidium bromide by spectroscopic method

Emodin interacting with deoxyribonucleic acid (DNA) has been studied by different spectroscopic techniques, such as fluorescence, ultraviolet and visible (UV-vis), and fourier transform infared (FT-IR) spectroscopies, using ethidium bromide (EB) as a fluorescence probe of DNA. The decrease in the fluorescence of DNA-EB system on addition of emodin shows that the fluorescence quenching of DNA-EB complex by emodin occurs. The binding constants of emodin with DNA in the presence of EB are 6.02x10(4), 9.20x10(4) and 1.17x10(5)Lmol(-1) at 20, 35 and 50 degrees C, respectively. FT-IR spectrum further suggests that both the phosphate groups and the bases of DNA react with emodin. The reaction of DNA with emodin in the presence of EB is affected by ionic strength and temperature. The values of melting temperature (T(m)) of DNA-EB complex and emodin-DNA-EB complexes were determined, respectively. From the experiment evidences, the major binding mode of emodin with DNA should be the groove binding.     

Complexation of DNA with cationic gemini surfactant in aqueous solution

Interactions between DNA and the cationic gemini surfactant trimethylene-1,3-bis(dodecyldimethylammonium bromide) (12-3-12) in aqueous solution have been investigated by UV-vis transmittance, zeta potential, and fluorescence emission spectrum. Complexes of DNA and gemini surfactant are observed in which the negative charges of DNA are neutralized by cationic surfactants effectively. The DNA-induced micelle-like structure of the surfactant due to the electrostatic and hydrophobic interactions is determined by the fluorescence spectrum of pyrene. It is found that the critical aggregation concentration (CAC) for DNA/12-3-12 complexes depends little on the addition of sodium bromide (NaBr) because of the counterbalance salt effect. However, at high surfactant concentration, NaBr facilitates the formation of larger DNA/surfactant aggregates. Displacement of ethidium bromide (EB) by surfactant evidently illustrates the strong cooperative binding between surfactant and DNA. In contrast to that in the absence of surfactant, the added NaBr at high surfactant concentration influences not only the binding of surfactant with DNA, but also the stability of DNA/EB complex.     

Spectroscopic studies of the interaction between pirimicarb and calf thymus DNA

The interaction between pirimicarb and calf thymus DNA in physiological buffer (pH 7.4) was investigated with the use of Neutral Red (NR) dye as a spectral probe by UV-vis absorption, fluorescence and circular dichroism (CD) spectroscopy, as well as viscosity measurements and DNA melting techniques. The results revealed that an intercalation binding should be the interaction mode of pirimicarb to DNA. CD spectra indicated that pirimicarb induced conformational changes of DNA. The binding constants of pirimicarb with DNA were obtained by the fluorescence quenching method. The thermodynamic parameters, enthalpy change (ΔHθ) and entropy change (ΔSθ) were calculated to be -52.13±2.04 kJ mol(-1) and -108.8±6.72 J mol(-1) K(-1) according to the van't Hoff equation, which suggested that hydrogen bonds and van der Waals forces might play a major role in the binding of pirimicarb to DNA. Further, the alternative least squares (ALS) method was applied to resolve a complex two-way array of the absorption spectra data, which provided simultaneously the concentration information for the three reaction components, pirimicarb, NR and DNA-NR. This ALS analysis indicated that the intercalation of pirimicarb into the DNA by substituting for NR in the DNA-NR complex.     

Accidental discovery of a ‘longer-range’ vinylogous Pummerer-type lactonization: formation of sulindac sulfide lactone from sulindac

Unexpected formation of sulindac sulfide lactone occurred when sulindac was treated with oxalyl chloride and triethylamine. Structurally analogous sulindac sulfide and indomethacin did not undergo such lactonization under similar reaction conditions. We believe that the sulfoxide function in sulindac plays a pivotal role possibly via a ‘longer-range’ vinylogous Pummerer-type reaction as a driving force for the observed lactonization. The structure of the lactone was confirmed by single crystal X-ray analysis.     

光谱法和电化学法研究甲硫氨酸二肽与DNA的相互作用

利用紫外-可见光谱、荧光探针、循环伏安等方法研究了甲硫氨酸二肽(Met-Met)与DNA的相互作用.结果发现:加入甲硫氨酸二肽后,DNA-Met-Met体系的紫外光谱呈减色效应,同时甲硫氨酸二肽的加入使得EB-DNA荧光强度减弱;循环伏安法表明,DNA的加入使得甲硫氨酸二肽的峰电流减小,峰位负移;Stern-Volmer方程说明二肽对DNA-EB的作用属于静态猝灭.这几种方法的实验结果都表明两者的作用模式为静电结合,多种计算方法得到两者作用的结合常数达到103 L/mol.