CN110498795B - 咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针及其应用 - Google Patents
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Abstract
本发明公开了一种咪唑并[1,2‑a]吡啶衍生物类半胱氨酸荧光探针，其化学结构式如式（1）所示。本发明的荧光探针在DMSO/水(体积比1:9)溶液中对半胱氨酸有较好的荧光选择性、较高的灵敏度及较强的抗其它离子干扰能力，具有巨大的应用前景。
Description
技术领域
本发明涉及有机小分子荧光探针领域，尤其涉及一种咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针的合成及其应用。
背景技术
半胱氨酸是一种具有生理功能,在自然界中广泛存在的重要物质,它在动植物体内是各组织细胞用来防御有害物质和增加活力的一种氨基酸,也是组成蛋白质的多种氨基酸之一,并且是唯一具有活性疏基的氨基酸。半胱氨酸是乙酞辅酶、牛磺酸和还原谷胱甘肽的前体,同时还能与铁离子形成硫铁化合物。目前,研究表明,人体中半胱氨酸浓度的升高是引起心血管疾病及中风的重要因素,还会导致认知功能障碍,严重的会导致阿尔茨海默氏病。因此,发明一种能方便检测生理环境中半胱氨酸的方法具有重要的现实意义。
目前常用的半胱氨酸的方法主要有以下几种,如电化学检测法、高效液相色谱法,分光光度法、电化学检测法,这些方法只能检测半胱氨酸的总量,并且样品需求量大、检测时间长,而且会对样品造成破坏,因此并不适于生物环境的半胱氨酸的检测。因此,探索方便廉价、能够定性定量检测半胱氨酸,并且可以对生物样品进行实时检测的方法非常重要。当有机荧光探针与特定目标物发生变化后,其荧光信号会发生变化,以达到检测的目的。荧光分析法具有灵敏度高、选择性好、取样量少、方法简洁快速等特征。所以,发明能快速检测、易观查信号变化的半胱氨酸荧光探针是非常有必要的。
发明内容
针对现有技术的不足，本发明解决的问题是提供一种咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针及其应用。
本发明的技术方案是：一种咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针，其化学结构式如式（1）所示：
本发明还包括咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针的应用，式（1）化合物在DMSO/水(体积比1:9)溶液中对半胱氨酸有独特的荧光选择性。
本发明还包括咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针的合成方法：在二氯甲烷溶液中，将2-(4-咪唑并[1,2-a]吡啶基)苯酚丙烯酰氯与三乙胺按照摩尔比1:2:3投料，加热回流4小时，得到咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针。
配制咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针的DMSO/水(体积比1:9)溶液，分别加入定量的Cys, Ala, Tyr, Trp, His, Val, Lle, Arg, Glu, Pry, Gly, LysAsp, Phe, Met, Ser, Leu的水溶液，通过荧光光谱测试来研究对不同氨基酸的选择性，测其荧光发射波谱强度变化发现：本发明所述咪唑并[1,2-a]吡啶类半胱氨酸荧光探针即式（1）化合物对半胱氨酸有独特的荧光选择性，如图1所示。逐渐加入半胱氨酸至20当量后，化合物1在376nm处荧光强度明显升高，如图2所示。因此，咪唑并[1,2-a]吡啶衍生物即式（1）化合物作为半胱氨酸荧光探针具有巨大的应用。
附图说明
图1：式（1）化合物（10-6M）的DMSO/水(体积比1:9)溶液中加入20当量的不同氨基酸后的荧光强度变化图，图1中FL Intensity为荧光强度。
图2：式（1）化合物（10-6M）的DMSO/水(体积比1:9)溶液中进行半胱氨酸荧光滴定图。图中FL Intensity为荧光强度，Wavelength为波长，equiv为倍数。
图3：式（1）化合物（10-6M）和20当量的半胱氨酸共存的DMSO/水(体积比1:9)溶液中加入20当量的其它氨基酸后荧光强度比率的变化柱状图, 图中FL Intensity为荧光强度。
图4是式（1）化合物合成方法反应式图。
具体实施方式
实施例1：式（1）化合物的合成方案如下式所示：
具体合成步骤如下：
在50 mL圆底烧瓶中依次加入0.2g（0.954 mmol）2-(4-咪唑并[1,2-a]吡啶基)苯酚，0.17g（1.9mmol）丙烯酰氯，0.73g三乙胺，15 mL二氯甲烷，加热回流4小时。TLC检测反应完成后，加入硅胶浓缩，柱层析得0.12g白色固体，产率48%。
核磁共振氢谱测定：1H NMR（400 MHz，CDCl3）：δ8.32 (m，1H),8.08 (d， 1H), 7.80(s, 1H), 7.63 (d，1H),7.38 (m, 2H),7.18 (m，2H) 6.77 (m, 1H), 6.68 (d, 1H),6.46 (m, 1H), 6.10 (d, 1H)。
实施例2：
向式（1）化合物（10-6M）的DMSO/水(体积比1:9)溶液中分别加入20当量的Cys,Ala, Tyr, Trp, His, Val, Lle, Arg, Glu, Pry, Gly, Lys Asp, Phe, Met, Ser, Leu后，测其376nm处荧光发射强度变化发现：式（1）化合物对半胱氨酸有独特的荧光选择性，加入20当量的半胱氨酸后，化合物1在376nm处荧光强度明显升高，如图1，2所示。
实施例3：
于式（1）化合物（10-6M）和20当量的半胱氨酸的DMSO/水(体积比1:9)溶液中分别加入等当量的Cys, Ala, Tyr, Trp, His, Val, Lle, Arg, Glu, Pry, Gly, Lys Asp,Phe, Met, Ser, Leu后，测其在376nm荧光发射强度变化发现：式（1）化合物对其它离子有较强的抗干扰能力，如图3所示。
Claims (2)
1.一种咪唑并[1,2-a]吡啶衍生物类半胱氨酸荧光探针，其特征在于:它的化学结构式如式(1)所示:
(1)
式(1)化合物在体积比为1:9的DMSO/水溶液中，对半胱氨酸有荧光选择性。
2.式(1)所述咪唑并[1.2-a]吡啶衍生物类半胱氨酸荧光探针的合成方法，其特征在于:在二氯甲烷溶液中，将式(2)化合物、丙烯酰氯与三乙胺按摩尔比 1:2:3 投料，加热回流4小时，得到权利 要求1中的式(1)化合物
(2) 。
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