Synthesis o novel arylboronic acids and their analytical use as luminescent reagents

新型芳基硼酸的合成及其作为发光试剂的分析用途

• 批准号: 10672021
• 负责人: KURODA Naotaka
• 金额: $ 1.98万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672021/
• 关键词: arylboronic acid derivatives; lophine; chemiluminescence; horseradish peroxidase; photographic detection; enhancer; fluorescence; アリールボロン酸; ペルオキシダーゼ; 糖; ジオール

Four kinds of novel arylboronic acids, which are the derivatives of lophine and its analogues, were synthesized based on the fact that lophine shows strong fluorescence (FL) and chemiluminescence (CL).Since some arylboronic acids are known to enhance the luminol-hydrogen peroxide-horseradish peroxidase (HRP) CL reaction, we evaluated those as enhancers at first. Among the compounds examined, 4-[4,5-di(2-pyridyl)-1H-imidazol-2-yl]phenyl boronic acid (DPPA) was found to be the most potent enhancer. The detection limit of HRP with DPPA was 0.15 ng (ca. 3.5 fmol) which was ca. 180 times better than that in the absence of the enhancer. By using DPPA with the effect of prolonging the light emission, the CL system was applied to the photographic detection of HRP on a nitrocellulose membrane. After optimizing the CL reaction conditions, as little as 0.1 ng of HRP was photographically detectable. This method was successfully applied to the western blotting assay of anti-human T-cell leukemia virus type-I.Next, FL and CL characteristics of the arylboronic acids were examined as fundamental studies on developing FL probes or FL/CL labeling reagents for diol compounds. 4-(4,5-Diphenyl-1H-imidazol-2-yl )phenylboronic acid (DPA) showed a stronger FL than lophine whereas the other derivatives had weaker FL. Stronger CL was not observed for all derivatives, compared to that of lophine. Although the reaction of DPA with some diols was estimated by TLC so far, further investigation on the reactivity and spectral studies will be required.

基于罗非碱具有强荧光和化学发光的特性，合成了4种新的芳基硼酸类化合物，它们是罗非碱及其类似物的衍生物，由于一些芳基硼酸类化合物对鲁米诺-过氧化氢-辣根过氧化物酶（HRP）化学发光反应具有增强作用，本文首先对其作为增强剂进行了评价。在所检测的化合物中，发现4-[4，5-二（2-吡啶基）-1H-咪唑-2-基]苯基硼酸（DPPA）是最有效的增强剂。DPPA对HRP的检测限为0.15 ng（约1.5 μ g）。3.5 fmol），其为ca.比没有增强剂的情况好180倍。利用DPPA的发光延长效应，将化学发光体系应用于硝酸纤维素膜上HRP的照相检测。在优化化学发光反应条件后，低至0.1 ng的HRP是可检测的。该方法成功地应用于抗人T细胞白血病病毒I型的蛋白质印迹分析。接下来，FL和CL的芳基硼酸的特性进行了检查，作为开发FL探针或FL/CL标记试剂的二醇化合物的基础研究。4-（4，5-二苯基-1H-咪唑-2-基）苯硼酸（DPA）的荧光强度比洛啡碱强，而其他衍生物的荧光强度较弱。虽然DPA与一些二元醇的反应迄今为止是通过TLC估计的，但需要进一步调查反应性和光谱研究。

项目成果

K.Nakashima: "Fluorescence and chemiluminescence of newly developed lophine analogues" Dyes and Pigments. 38. 127-136 (1998)

K.Nakashima：“新开发的洛芬类似物的荧光和化学发光”染料和颜料。

N. Kuroda, et al.: "Photographic detection of fluorescent-labelled oligodeoxynucleotide in the blotting format by peroxyoxalate chemiluminescence"J. Biolumin. Chemilumin.. 13. 101-105 (1998)

N. Kuroda 等人：“通过过氧草酸盐化学发光以印迹形式对荧光标记的寡脱氧核苷酸进行照相检测”。

K.Nakashima,et al: "Fluorescence and chemiluminescence of mewlydeveloped lophine analogues"Dyes and Pigments. 136. 127-136 (1998)

K.Nakashima 等人：“新开发的洛芬类似物的荧光和化学发光”染料和颜料。

N.Kuroda, et al.: "New phenylboronic acid derivatives as enhancers for the luminol-HィイD22ィエD2OィイD22ィエD2-horseradish peroxidase chemiluminescence reaction"Luminescence. 14. 361-364 (1999)

N. Kuroda 等人：“新的苯基硼酸衍生物作为鲁米诺-D22D2OD22-辣根过氧化物酶化学发光反应的增强剂”，发光，14. 361-364 (1999)。

N.Kuroda: "Chemiluminescent derivatization of adenyl compounds with glyoxal derivatives in the presence of heteropoly acids and its application to the simple and sensitive determination of DNA" J.Biolumin.Chemilumin.13. 25-29 (1998)

N.Kuroda：“在杂多酸存在下，腺苷酸化合物与乙二醛衍生物的化学发光衍生化及其在简单灵敏的 DNA 测定中的应用”J.Biolumin.Chemilumin.13。