CN110041317B - 一种萘酰亚胺荧光探针及其制备与应用 - Google Patents
一种萘酰亚胺荧光探针及其制备与应用 Download PDFInfo
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Abstract
本发明公开了一种基于萘酰亚胺的荧光探针及其制备方法和应用。其是在1,8‑萘酰亚胺的荧光骨架中引入2‑噻吩甲酰基作为识别基团构建的。2‑噻吩甲酰基的引入，抑制了荧光团的ICT效应，因而荧光很弱。与小分子生物硫醇反应后，探针释放出强荧光的羟基萘酰亚胺，光物理性质改变，具有较大的Stokes位移，从而可作为生物硫醇检测的荧光探针。又通过反应动力学，计算了探针对谷胱甘肽、半胱氨酸、高半胱氨酸的伪一级反应速率和二级反应速率，借此对这三种性质相近的生物硫醇加以区分。实验证明，该探针对GSH和Cys表现出高灵敏度和高选择性，且具有较低的检测限，可用于斑马鱼体内的生物硫醇荧光成像检测。
Description
技术领域
本发明属于分析化学领域，具体涉及一种2-噻吩甲酰基修饰的萘酰亚胺荧光探针及其制备方法与其在生物硫醇的荧光检测方面的应用。
背景技术
小分子硫醇在维持细胞氧化还原环境和减轻自由基和毒素损害方面发挥着至关重要的作用[G.I.Giles. The redox regulation of thiol dependent signalingpathways in cancer. Current pharmaceutical design, 2006, 12(34): 4427-4443；N.Brandes, S. Schmitt, U.Jakob. Thiol-based redox switches in eukaryoticproteins. Antioxidants & redox signaling, 2009, 11(5): 997-1014.]。生物系统中常见三种低分子量硫醇，即半胱氨酸（Cys），高半胱氨酸（Hcy）和谷胱甘肽（GSH）。这些硫醇密切参与调节各种生理和病理过程。例如，Cys是蛋白质合成的必需氨基酸，其异常水平将导致增长缓慢、浮肿、嗜睡、肝功能损害等[S.Shahrokhian. Lead phthalocyanine as aselective carrier for preparation of a cysteine-selective electrode.Analytical Chemistry, 2001, 73(24): 5972-5978；M.T.Heafield, S.Fearn,G.B.Steventon, R.H. Waring, A.C. Williams, S.G.Sturman. Plasma cysteine andsulphate levels in patients with motor neurone, Parkinson's and Alzheimer'sdisease. Neuroscience Letters, 1990, 110(1-2): 216-220.]。另一方面，血浆中Hcy升高是心血管疾病、阿尔茨海默氏病和骨质疏松症的危险因素[P Sachdev, R.Parslow,C.Salonikas, O. Lux, W. Wen, R.KμMar, D. Naidoo, H. Christensen, A Jorm.Homocysteine and the brain in midadult life: evidence for an increased riskof leukoaraiosis in men. Archives of Neurology, 2004, 61(9): 1369-1376.]。而GSH则与白细胞减少、癌症、HIV感染等紧密相连[S.C. Lu. Regulation of glutathionesynthesis. Molecular Aspects of Medicine, 2009, 30(1-2): 42-59; D.M.Townsend, K.D.Tew, H.Tapiero. The importance of glutathione in human disease.Biomedicine & Pharmacotherapy, 2003, 57(3-4): 145-155.]。硫醇的重要生物作用促使研究人员对开发有用的化学工具用于检测硫醇产生浓厚的兴趣。
荧光探针由于操作简单、对生物体损害小已成为活细胞中硫醇检测的常用方法。在过去十年中，已开发出大量用于检测硫醇的荧光探针。这些探针大多数是基于硫醇的选择性化学反应构建的，包括迈克尔加成反应[M.JungáKim.A thiol-specific fluorescentprobe and its application for bioimaging. Chemical Communications, 2010, 46(16): 2751-2753; H. Zhang, P. Wang, Y.X. Yang, H.Y. Sun. A selectivefluorescent probe for thiols based on α, β-unsaturated acyl sulfonamide.Chemical Communications, 2012, 48(86): 10672-10674.]，亲核取代[D. Kand, A.M.Kalle, S.J. Varma, P. Talukdar. A chromenoquinoline-based fluorescent off–onthiol probe for bioimaging. Chemical Communications, 2012, 48(21): 2722-2724;J. Liu, Y.Q. Sun, H. Zhang, Y.Y. Hou, Y.W. Shi, W. Guo. Simultaneousfluorescent imaging of Cys/Hcy and GSH from different emission channels.Chemical Science, 2014, 5(8): 3183-3188.]，醛和氨基硫醇之间环化反应[H.L. Li,J.L. Fan, J.Y. Wang, M.A. Tian, J.J. Du, S.G. Sun, P.P. Sun, X.J. Peng. Afluorescent chemodosimeter specific for cysteine: effective discrimination ofcysteine from homocysteine. Chemical Communications, 2009 (39): 5904-5906;Z.G. Yang, N. Zhao, Y.M. Sun, F. Miao, Y. Liu, X. Liu, Y.H. Zhang, W.T. Ai,G.F. Song, X.Y. Shen, X.Q. Yu, J.Z. Sun, W.Y. Wong. Highly selective red-andgreen-emitting two-photon fluorescent probes for cysteine detection and theirbio-imaging in living cells. Chemical Communications, 2012, 48(28): 3442-3444.]，2,4-二硝基苯（DNBS）与硫醇的裂解反应[J. Bouffard, Y. Kim, T.M. Swager,R. Weissleder, S.A. Hilderbrand. A highly selective fluorescent probe forthiol bioimaging. Organic letters, 2008, 10(1): 37-40.]，二硫化物交换反应[C.S.Lim, G. Masanta, H.J. Kim, J.H. Han, H.M. Kim, B.R. Cho. Ratiometricdetection of mitochondrial thiols with a two-photon fluorescent probe.Journal of the American Chemical Society, 2011, 133(29): 11132-11135.]等等。
本发明通过对1.8-萘二甲酰亚胺的结构进行改造，合成了一种2-噻吩甲酰基修饰的萘酰亚胺荧光探针，并实现了其在生物硫醇荧光检测中的应用，具有良好的发展前景。
发明内容
本发明的目的在于提供一种2-噻吩甲酰基修饰的萘酰亚胺荧光探针及其制备方法与其在生物硫醇荧光检测中的应用。
为了实现上述目的，本发明采用如下的技术方案：
一种2-噻吩甲酰基修饰的萘酰亚胺荧光探针，其结构式如下：
所述2-噻吩甲酰基修饰的萘酰亚胺荧光探针的制备方法包括以下步骤：
（1）将羟基取代的萘酰亚胺衍生物
和三乙胺在冰浴条件下混合,溶于溶剂，再逐滴滴入2-噻吩甲酰氯
，三者摩尔比1:1.2:2，滴加完毕后撤去冰浴，室温反应2个小时；
（2）反应完成后，减压除去溶剂，得粗产品；
（3）将粗产品经硅胶柱层析，得到所述2-噻吩甲酰基修饰的萘酰亚胺荧光染料。
其中，所述溶剂为二氯甲烷；
所述硅胶柱层析采用体积比为15:1的石油醚和乙酸乙酯的混合溶液做洗脱剂。
进一步地，所述羟基取代的萘酰亚胺衍生物的制备方法包括以下步骤：
（1）将甲氧基取代的萘酰亚胺衍生物
和47wt%的氢碘酸按5.5 mg/mL的质量体积比混合，回流反应过夜；
（2）反应结束后，冷却并减压抽滤，将收集的固体用水洗涤 (5 mL×3) 并真空干燥，得到粗产物；
（3）粗产物经硅胶柱层析得到所述羟基取代的萘酰亚胺衍生物。
其中，所述回流温度为127℃，回流时间为12小时；
所述硅胶柱层析采用体积比为20:1的石油醚和乙酸乙酯的混合溶液做洗脱剂。
进一步地，所述甲氧基取代的萘酰亚胺衍生物的制备方法包括以下步骤：
（1）将五水硫酸铜与正丁基取代的萘酰亚胺衍生物
和甲醇钠按摩尔比1:10:100混合于溶剂，回流反应过夜；
（2）反应结束后，减压除去溶剂，得到粗产物；
（3）将粗产物经硅胶柱层析得到所述甲氧基取代的萘酰亚胺衍生物。
其中，所述溶剂为甲醇，回流温度为65℃，回流时间为12小时；
所述硅胶柱层析是采用体积比为100:1的二氯甲烷和甲醇的混合溶液做洗脱剂。
进一步地，所述正丁基取代的萘酰亚胺衍生物的制备方法包括以下步骤：
（1）将4-溴-1,8-萘二甲酸酐
与正丁胺按摩尔比1:4溶于溶剂，回流反应过夜；
（2）反应结束后减压除去溶剂，得到粗产物；
（3）将粗产物经硅胶柱层析得到所述正丁胺取代的萘酰亚胺衍生物。
其中，所述溶剂为乙醇，回流温度为78℃，回流时间为12小时；
所述硅胶柱层析是采用体积比为20:1的石油醚和乙酸乙酯的混合溶液为洗脱剂。
所得2-噻吩甲酰基修饰的萘酰亚胺荧光探针应用于荧光响应的生物硫醇检测。
本发明将2-噻吩甲酰基做为识别基团连接在萘酰亚胺荧光团上。通过生物硫醇选择性地剪去探针分子上具有减弱荧光团ICT效应的2-噻吩甲酰基，使荧光团上的羟基得以释放，从而增强荧光团的ICT效应，起到改变探针光物理性质的作用，实现对小分子生物硫醇的定量测量。
本发明所得萘酰亚胺荧光探针对小分子生物硫醇Cys、GSH、Hcy具有良好的灵敏度和选择性，检测限分别达到0.065 μM，0.120 μM和0.339 μM。同时，通过动力学研究表明，探针对它们的响应速率为Cys＞GSH＞Hcy，该工作为具有ICT效应的生物硫醇荧光探针的设计提供了新的方案。
附图说明
图1为萘酰亚胺荧光探针（10 μM）及加入GSH（50 μM）后分别在不同pH（3-9）的缓冲溶液中的荧光响应图。结果显示为三次单独测量的平均值±标准差。(λex =450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图2为在PBS缓冲溶液（10 mM, pH 7.4, 1 mM CTAB）中，萘酰亚胺荧光探针（10 μM）在加入GSH（50 μM）后分别在不同温度（25-45℃）下孵育2 h的荧光响应图。结果显示为三次单独测量的平均值±标准差。(λex=450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图3为萘酰亚胺荧光探针（10 μM）滴加不同浓度GSH（0-200 μM）的荧光响应光谱图。(λex =450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图4为萘酰亚胺荧光探针（10 μM）在550 nm处的荧光强度与GSH浓度（5-30 μM）之间的线性拟合曲线。(λex =450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图5为萘酰亚胺荧光探针（10 μM）滴加不同浓度Cys（0-200 μM）的荧光响应光谱图。(λex =450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图6为萘酰亚胺荧光探针（10 μM）在550 nm处的荧光强度与Cys浓度（0-30 μM）之间的线性拟合曲线。(λex =450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图7为萘酰亚胺荧光探针（10 μM）滴加不同浓度Hcy（0-300 μM）的荧光响应光谱图。(λex =450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图8为萘酰亚胺荧光探针（10 μM）在550 nm处的荧光强度与Hcy浓度（5-50 μM）之间的线性拟合曲线。(λex =450 nm, slit:10 nm/10 nm, PMT Volts: 450 V)。
图9斑马鱼的共聚焦荧光成像图。（a）（b）（c）只用萘酰亚胺荧光探针（10 μM）孵育20分钟，（d）（e）（f）先用N-乙基马来酰亚胺（NEM, 200 μM）孵育30分钟，然后用萘酰亚胺荧光探针（10 μM）孵育20分钟。（a）（d）明场与暗场叠加，（b）（e）暗场，（c）（f）明场。
具体实施方式
为了使本发明所述的内容更加便于理解，下面结合具体实施方式对本发明所述的技术方案做进一步的说明，但是本发明不仅限于此。
实施例1
正丁基取代的萘酰亚胺衍生物
的制备：
在100 mL圆底烧瓶中，将4-溴-1,8-萘二甲酸酐（0.84 g, 3.70 mmol）和正丁胺（1.46 mL, 1.08 g, 14.8 mmol）溶于50 mL乙醇中，回流过夜。反应结束后，减压除去溶剂乙醇，硅胶柱提纯，洗脱剂为PE:EA=20:1(v/v)，得到白色固体产物775 mg，产率63%。1H NMR(400 MHz, CDCl3) δ 8.65 (d, J = 7.2 Hz, 1H), 8.56 (d, J = 8.5 Hz, 1H), 8.41(d, J = 7.9 Hz, 1H), 8.04 (d, J = 7.8 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 4.17(t, J = 7.5 Hz, 2H), 1.75-1.68 (m, 2H), 1.50-1.40 (m, 2H), 0.98 (t, J = 7.3Hz, 3H); 13C NMR (100 MHz, CDCl3) δ 163.44, 163.41, 133.00, 131.85, 131.04,130.97, 130.41, 130.03, 128.78, 127.96, 123.03, 122.16, 40.35, 30.14, 20.37,13.84; HRMS (ESI): Calcd for C16H15NO2Br ([M+H]+): 332.0286, Found: 332.0290.
实施例2
甲氧基取代的萘酰亚胺衍生物
的制备：
于100 mL圆底烧瓶中，实施例1中制备的正丁基取代的萘酰亚胺衍生物（1.49 g,4.50 mmol），甲醇钠（2.43 g, 45.0 mmol）和五水合硫酸铜（112 mg, 0.45 mmol）溶于40mL甲醇中，回流过夜。反应结束后，减压除去溶剂甲醇，硅胶柱提纯，洗脱剂为DCM:MeOH=100:1(v/v)，得到浅黄色固体产物956 mg，产率75%。1H NMR (400 MHz, CDCl3) δ 8.57 (d,J = 7.3 Hz, 1H), 8.52 (d, J = 8.3 Hz, 2H), 7.67 (t, J = 7.8 Hz, 1H), 7.01 (d,J = 8.3 Hz, 1H), 4.16 (t, J = 7.5 Hz, 2H), 4.12 (s, 3H), 1.75-1.68 (m, 2H),1.50-1.40 (m, 2H), 0.98 (t, J = 7.4 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ163.72, 163.10, 159.98, 132.55, 130.64, 128.39, 127.73, 125.25, 122.57,121.67, 114.40, 104.62, 55.84, 39.72, 30.05, 20.28, 13.74; HRMS (ESI): Calcdfor C17H18NO3 ([M+H]+): 284.1287, Found: 284.1289.
实施例3
羟基取代的萘酰亚胺衍生物
的制备：
于50 mL圆底烧瓶中，将实施例2中制备的甲氧基取代的萘酰亚胺衍生物（110 mg,0.39 mmol）和20 mL 47wt%氢碘酸的混合物回流12小时。反应结束后，冷却并减压抽滤，将收集的固体用水洗涤 (5 mL×3) 并真空干燥，硅胶柱提纯，洗脱剂为PE:EA=20:1(v/v)，得到黄色针状固体产物93 mg，产率89%。1H NMR (400 MHz, d 6-DMSO) δ 8.52 (d, J = 8.3Hz, 1H), 8.45 (d, J = 7.2 Hz, 1H), 8.35 (d, J = 8.2 Hz, 1H), 7.75 (t, J = 7.8Hz, 1H), 7.15 (d, J = 8.2 Hz, 1H), 4.02 (t, J = 7.4 Hz, 2H), 1.64-1.56 (m,2H), 1.39-1.30 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H); 13C NMR (100 MHz, d 6-DMSO) δ164.12, 163.45, 160.69, 133.99, 131.56, 129.60, 129.32, 126.04, 122.83,122.26, 113.07, 110.41, 30.21, 20.29, 14.19; HRMS (ESI): Calcd for C16H16NO3([M+H]+): 270.1130, Found: 270.1129.
实施例4
2-噻吩甲酰基修饰的萘酰亚胺荧光探针
的制备：
于25 mL圆底烧瓶中，在冰浴条件下将实施例3中制备的羟基取代的萘酰亚胺衍生物（67 mg, 0.25 mmol）和干燥的三乙胺（30 mg, 0.30 mmol）溶于5 mL干燥的二氯甲烷中，之后逐滴滴入2-噻吩甲酰氯（73 mg, 0.50 mmol），撤去冰浴，室温反应2小时，反应结束后旋干溶剂，硅胶柱层析，洗脱剂为PE:EA=15:1(v/v)，得到白色固体产物72 mg，产率76%。1HNMR (400 MHz, CDCl3) δ 8.67 (dd, J = 7.6, 3.9 Hz, 2H), 8.37 (d, J = 8.4 Hz,1H), 8.16 (d, J = 3.4 Hz, 1H), 7.81 (t, J = 7.6 Hz 2H), 7.74 (d, J = 8.0 Hz,1H), 7.30 (t, J =4.9 Hz, 1H), 4.22 (t, J =7.5 Hz, 2H), 1.79-1.72 (m, 2H),1.53-1.44 (m, 2H), 1.01 (t, J = 7.3 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ164.07, 163.52, 159.68, 151.29, 135.71, 134.66, 131.76, 131.72, 131.57,129.39, 128.53, 127.74, 127.35, 125.36, 123.03, 120.62, 119.54, 40.34, 30.22,20.40, 13.86; HRMS (ESI): Calcd for C21H18NO4S ([M+H]+): 380.0957, Found:380.0951.
应用实施例
1.生理环境下，pH也会有一定范围的浮动。为了研究探针在复杂生理环境中的适用性，研究了在不同pH值（3、4、5、6、7、7.4、8、9）的PBS缓冲溶液中的荧光响应变化，结果见图1。
由图1 可知，在pH 3.0-6.0范围内，萘酰亚胺荧光探针在550 nm处都处于无荧光状态，而在pH 6.0-9.0范围内，萘酰亚胺荧光探针在550 nm处的荧光有微弱的增强，但总体来说没有明显的变化。说明萘酰亚胺荧光探针在pH 3.0-9.0范围内有良好的稳定性。向含有萘酰亚胺荧光探针的反应体系中加入GSH后，在pH 3.0-5.0范围内550 nm处的荧光强度没有明显变化，而在pH 5.0-9.0范围内荧光强度有显著的增强，且在pH 7.0-9.0范围内基本持平。实验结果表明，萘酰亚胺荧光探针能够应用于生理环境下GSH 的检测。在后续的实验中选择PBS缓冲液pH 7.4作为测试pH。
2.将只含有萘酰亚胺荧光探针及加入GSH（50 μM）后的PBS缓冲溶液（10 mM, pH7.4, 1 mM CTAB）分别于25℃、30℃、35℃、40℃、45℃恒温摇床中孵育2小时，检测其在550nm处的荧光强度，结果见图2。
由图2可知，萘酰亚胺荧光探针在不同温度下550 nm处的荧光强度都处于猝灭状态，且保持稳定。加入GSH后，荧光强度均有明显增强，且差别不明显。说明探针在该温度范围内有良好的稳定性。
3. 在含有10 μM实施例4所得萘酰亚胺荧光探针的PBS缓冲溶液（10 mM, pH 7.4,1 mM CTAB）中，进行GSH的荧光滴定实验，结果见图3、4。
由图3可见，在PBS缓冲溶液（10 mM, pH 7.4, 1 mM CTAB）中，550 nm处萘酰亚胺荧光探针的荧光处于猝灭状态，当向体系中加入GSH时，荧光强度逐渐增强，当加入GSH的浓度为50 μM时，荧光强度基本达到饱和，此时相对于未添加时的荧光强度有约20倍的增强。
如图4所示，GSH浓度在5-30 μM范围内，溶液的荧光强度与GSH浓度呈现良好的线性关系（R2=0.987）。根据公式3σ/k，计算得其检测限为0.120 μM。
4.在含有10 μM实施例4所得萘酰亚胺荧光探针的PBS缓冲溶液（10 mM, pH 7.4,1 mM CTAB）中，进行Cys的荧光滴定实验，结果见图5、6。
由图5可见，在PBS缓冲溶液（10 mM, pH 7.4, 1 mM CTAB）中，550 nm处萘酰亚胺荧光探针的荧光处于猝灭状态，当向体系中加入Cys时，荧光强度逐渐增强，当加入Cys的浓度为50 μM时，荧光强度基本达到饱和，此时相对于未添加时的荧光强度有约20倍的增强。
如图6所示，Cys浓度在0-30 μM范围内，溶液的荧光强度与Cys浓度呈现良好的线性关系（R2=0.992）。根据公式3σ/k，计算得其检测限为0.065 μM。
5.在含有10 μM实施例4所得萘酰亚胺荧光探针的PBS缓冲溶液（10 mM, pH 7.4,1 mM CTAB）中，进行Hcy的荧光滴定实验，结果见图7、8。
由图7可见，在PBS缓冲溶液（10 mM, pH 7.4, 1 mM CTAB）中，550 nm处萘酰亚胺荧光探针的荧光处于猝灭状态，当向体系中加入Hcy时，荧光强度逐渐增强，当加入Hcy的浓度为200 μM时，荧光强度基本达到饱和，此时相对于未添加时的荧光强度有约20倍的增强。
如图8所示，Hcy浓度在0-30 μM范围内，溶液的荧光强度与Hcy浓度呈现良好的线性关系（R2=0.993）。根据公式3σ/k，计算得其检测限为0.339 μM。
6.为了进一步证明所述的萘酰亚胺荧光探针可应用于生物体内成像，我们探究了其在斑马鱼体内检测生物硫醇的应用。选取经PTU处理的5天龄斑马鱼幼鱼用探针（10 μM）孵育30分钟，然后将其用PBS溶液冲洗后，在荧光共聚焦显微镜下进行荧光成像，拍摄得到图9中的(d) (e) (f)，可以观察到斑马鱼体内有出现黄色荧光。作为对照，我们将另一组斑马鱼幼鱼先用N-乙基马来酰亚胺（NEM, 200 μM）孵育20分钟，消耗掉其体内生物硫醇，再用探针（10 μM）孵育30 分钟，然后进行荧光共聚焦显微成像，结果看不到黄色荧光，如图9中的(a) (b) (c)。两组实验结果表明，我们可以通过该萘酰亚胺荧光探针检测到斑马鱼体内的硫醇，并进行体内成像研究。
以上所述仅为本发明的较佳实施例，凡依本发明申请专利范围所做的均等变化与修饰，皆应属本发明的涵盖范围。
Claims (7)
1.一种2-噻吩甲酰基修饰的萘酰亚胺荧光探针，其特征在于：其结构式如下：
2.一种如权利要求1所述的2-噻吩甲酰基修饰的萘酰亚胺荧光探针的制备方法，其特征在于：包括以下步骤：
（1）将羟基取代的萘酰亚胺衍生物
和三乙胺在冰浴条件下混合溶于溶剂，再逐滴滴入2-噻吩甲酰氯
，撤去冰浴，室温反应2个小时；
（2）反应完成后减压除去溶剂，得粗产品；
（3）粗产品经硅胶柱层析，得到所述2-噻吩甲酰基修饰的萘酰亚胺荧光染料；
其中，所述溶剂为二氯甲烷；
所述硅胶柱层析采用体积比为15:1的石油醚和乙酸乙酯的混合溶液做洗脱剂。
3.根据权利要求2所述的制备方法，其特征在于：所述羟基取代的萘酰亚胺衍生物的制备方法包括以下步骤：
（1）将甲氧基取代的萘酰亚胺衍生物
和47wt%的氢碘酸混合回流反应；
（2）反应结束后，冷却并减压抽滤，将收集的固体用水洗涤并真空干燥，得到粗产物；
（3）粗产物经硅胶柱层析得到所述羟基取代的萘酰亚胺衍生物；
其中，所述回流温度为127℃，回流时间为12小时；
所述硅胶柱层析采用体积比为20:1的石油醚和乙酸乙酯做洗脱剂。
4.根据权利要求3所述的制备方法，其特征在于：所述甲氧基取代的萘酰亚胺衍生物的制备方法包括以下步骤：
（1）将正丁基取代的萘酰亚胺衍生物
和甲醇钠、五水硫酸铜混合于溶剂，回流反应过夜；
（2）反应结束后，减压除去溶剂，得到粗产物；
（3）将粗产物经硅胶柱层析得到所述甲氧基取代的萘酰亚胺衍生物；
其中，所述溶剂为甲醇，回流温度为65℃，回流时间为12小时；
所述硅胶柱层析是采用体积比为100:1的二氯甲烷和甲醇的混合溶液做洗脱剂。
5.根据权利要求4所述的制备方法，其特征在于，正丁基取代的萘酰亚胺衍生物的制备方法包括以下步骤：
（1）将4-溴-1,8-萘二甲酸酐
与正丁胺溶于溶剂，回流反应过夜；
（2）反应结束后减压除去溶剂，得到粗产物；
（3）将粗产物经硅胶柱层析得到所述正丁基取代的萘酰亚胺衍生物；
其中，所述溶剂为乙醇，回流温度为78℃，回流时间为12小时；
所述硅胶柱层析是采用体积比为20:1的石油醚和乙酸乙酯混合溶液做洗脱剂。
6.一种如权利要求1所述的2-噻吩甲酰基修饰的萘酰亚胺荧光探针在生物硫醇检测中的应用，其特征在于：将2-噻吩甲酰基修饰的萘酰亚胺制成荧光增强响应的生物硫醇探针，并借助动力学差异，对三种性质相近的生物硫醇谷胱甘肽、半胱氨酸、高半胱氨酸加以区分。
7.根据权利要求6所述的应用，其特征在于：2-噻吩甲酰基修饰的萘酰亚胺荧光探针用于斑马鱼体内生物硫醇的荧光成像检测。
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673278A (zh) * | 2015-02-15 | 2015-06-03 | 浙江理工大学 | 一种检测谷胱甘肽的荧光探针及其制备方法与使用方法 |
| CN104710979A (zh) * | 2015-02-15 | 2015-06-17 | 浙江理工大学 | 一种用于检测谷胱甘肽的荧光探针及其制备方法与应用 |
| CN104797560A (zh) * | 2012-11-12 | 2015-07-22 | Az电子材料（卢森堡）有限公司 | 芳香族酰亚胺化合物及其制造方法 |
| CN105001161A (zh) * | 2015-06-08 | 2015-10-28 | 华中师范大学 | 一类基于1,8-萘酰亚胺为骨架的磺酰胺化合物、制备方法及其应用 |
| CN106929006A (zh) * | 2017-01-22 | 2017-07-07 | 浙江工业大学 | 一种以萘酰亚胺为母核的识别半胱氨酸和同型半胱氨酸荧光探针及其制备与应用 |
-
2019
- 2019-05-16 CN CN201910408358.0A patent/CN110041317B/zh not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104797560A (zh) * | 2012-11-12 | 2015-07-22 | Az电子材料（卢森堡）有限公司 | 芳香族酰亚胺化合物及其制造方法 |
| CN104673278A (zh) * | 2015-02-15 | 2015-06-03 | 浙江理工大学 | 一种检测谷胱甘肽的荧光探针及其制备方法与使用方法 |
| CN104710979A (zh) * | 2015-02-15 | 2015-06-17 | 浙江理工大学 | 一种用于检测谷胱甘肽的荧光探针及其制备方法与应用 |
| CN105001161A (zh) * | 2015-06-08 | 2015-10-28 | 华中师范大学 | 一类基于1,8-萘酰亚胺为骨架的磺酰胺化合物、制备方法及其应用 |
| CN106929006A (zh) * | 2017-01-22 | 2017-07-07 | 浙江工业大学 | 一种以萘酰亚胺为母核的识别半胱氨酸和同型半胱氨酸荧光探针及其制备与应用 |
Non-Patent Citations (2)
| Title |
|---|
| A fast response and high sensitivity thiol fluorescent probe in living cells;Tao Liu，等;《Sensors and Actuators B》;20160405;第232卷;第619-624页 * |
| Design and synthesis of a fluorescent probe based on naphthalene anhydride and its detection of copper ions;Guangjie He，等;《PLOS ONE》;20171026;第12卷(第10期);第1-11页 * |
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