CN102603733A - 3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉及其制法和在锌离子荧光传感与细胞成像中的应用 - Google Patents
3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉及其制法和在锌离子荧光传感与细胞成像中的应用 Download PDFInfo
- Publication number
- CN102603733A CN102603733A CN2011103599923A CN201110359992A CN102603733A CN 102603733 A CN102603733 A CN 102603733A CN 2011103599923 A CN2011103599923 A CN 2011103599923A CN 201110359992 A CN201110359992 A CN 201110359992A CN 102603733 A CN102603733 A CN 102603733A
- Authority
- CN
- China
- Prior art keywords
- phenanthroline
- pht1
- thienyl
- methyl
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000003384 imaging method Methods 0.000 title abstract description 13
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 title abstract description 10
- 238000002360 preparation method Methods 0.000 title abstract description 3
- KASZUPYSDPKHGH-UHFFFAOYSA-N 3,8-bis(5-methylthiophen-2-yl)-1,10-phenanthroline Chemical compound CC1=CC=C(S1)C=1C=NC2=C3N=CC(=CC3=CC=C2C1)C=1SC(=CC1)C KASZUPYSDPKHGH-UHFFFAOYSA-N 0.000 title abstract 2
- 108091006597 SLC15A4 Proteins 0.000 claims description 34
- 102100021484 Solute carrier family 15 member 4 Human genes 0.000 claims description 34
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical class C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 12
- 238000006243 chemical reaction Methods 0.000 claims description 9
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 claims description 8
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- 239000011777 magnesium Substances 0.000 claims description 6
- 238000010992 reflux Methods 0.000 claims description 6
- 229920006395 saturated elastomer Polymers 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 4
- LVEYOSJUKRVCCF-UHFFFAOYSA-N 1,3-Bis(diphenylphosphino)propane Substances C=1C=CC=CC=1P(C=1C=CC=CC=1)CCCP(C=1C=CC=CC=1)C1=CC=CC=C1 LVEYOSJUKRVCCF-UHFFFAOYSA-N 0.000 claims description 3
- GWKGPQCKIBRXGW-UHFFFAOYSA-N 5-bromo-1,10-phenanthroline Chemical compound C1=CC=C2C(Br)=CC3=CC=CN=C3C2=N1 GWKGPQCKIBRXGW-UHFFFAOYSA-N 0.000 claims description 3
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims description 3
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 3
- 229910052786 argon Inorganic materials 0.000 claims description 3
- 238000009835 boiling Methods 0.000 claims description 3
- 230000003197 catalytic effect Effects 0.000 claims description 3
- 239000003480 eluent Substances 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 229910052740 iodine Inorganic materials 0.000 claims description 3
- 239000011630 iodine Substances 0.000 claims description 3
- 229910052749 magnesium Inorganic materials 0.000 claims description 3
- 239000012044 organic layer Substances 0.000 claims description 3
- 229910002027 silica gel Inorganic materials 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 3
- 229960001866 silicon dioxide Drugs 0.000 claims description 3
- 235000002639 sodium chloride Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 230000000977 initiatory effect Effects 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims 1
- 239000011701 zinc Substances 0.000 abstract description 30
- 238000002474 experimental method Methods 0.000 abstract description 7
- 238000001514 detection method Methods 0.000 abstract description 6
- 229910001428 transition metal ion Inorganic materials 0.000 abstract description 4
- 229910001413 alkali metal ion Inorganic materials 0.000 abstract description 3
- 229910001420 alkaline earth metal ion Inorganic materials 0.000 abstract description 3
- 150000005045 1,10-phenanthrolines Chemical class 0.000 abstract description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 abstract description 2
- 230000007541 cellular toxicity Effects 0.000 abstract 1
- 239000011259 mixed solution Substances 0.000 abstract 1
- 210000000056 organ Anatomy 0.000 abstract 1
- 230000035699 permeability Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 24
- 239000000243 solution Substances 0.000 description 20
- 150000002500 ions Chemical class 0.000 description 18
- 229910052751 metal Inorganic materials 0.000 description 7
- 239000002184 metal Substances 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 229910021645 metal ion Inorganic materials 0.000 description 5
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 4
- 239000012981 Hank's balanced salt solution Substances 0.000 description 4
- 108091006595 SLC15A3 Proteins 0.000 description 4
- 102100021485 Solute carrier family 15 member 3 Human genes 0.000 description 4
- YTPLMLYBLZKORZ-UHFFFAOYSA-N Thiophene Chemical compound C=1C=CSC=1 YTPLMLYBLZKORZ-UHFFFAOYSA-N 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- YNPNZTXNASCQKK-UHFFFAOYSA-N Phenanthrene Natural products C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- -1 metals ion Chemical class 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 241000228143 Penicillium Species 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 description 2
- 230000002860 competitive effect Effects 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 239000004744 fabric Substances 0.000 description 2
- 230000012447 hatching Effects 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- QTENRWWVYAAPBI-YCRXJPFRSA-N streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O QTENRWWVYAAPBI-YCRXJPFRSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229930192474 thiophene Natural products 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 206010000830 Acute leukaemia Diseases 0.000 description 1
- 235000000621 Bidens tripartita Nutrition 0.000 description 1
- 240000004082 Bidens tripartita Species 0.000 description 1
- 101100184147 Caenorhabditis elegans mix-1 gene Proteins 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- JJWSNOOGIUMOEE-UHFFFAOYSA-N Monomethylmercury Chemical compound [Hg]C JJWSNOOGIUMOEE-UHFFFAOYSA-N 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 230000010748 Photoabsorption Effects 0.000 description 1
- 238000002441 X-ray diffraction Methods 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000000119 electrospray ionisation mass spectrum Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 208000006637 fused teeth Diseases 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003387 muscular Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000003352 sequestering agent Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000003335 steric effect Effects 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Images
Landscapes
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
一种1,10-菲罗啉衍生物,它是3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉(PHT1),具有如下结构式:。它在乙醇-水混合溶液中,能选择性螯合游离锌离子,形成化学计量比2:1的锌离子配合物([Zn(PHT1)2]2+),配合物稳定常数高达3.4×1012M-1,且在蓝光区域(455~490nm)产生51倍的荧光增强。这种荧光增强既不受碱金属离子(如Na+,K+),碱土金属离子(如Ca2+,Mg2+)以及大部分过渡金属离子(如Hg2+,Pb2+,Ag+)等影响,也不受pH影响,对游离锌离子的检测限可以达到5ppb。HL-60细胞、HepG-2细胞的成像实验表明PHT1具有良好的细胞渗透性以及低的细胞毒性,而且能够标记细胞器中痕量的锌离子,能应用于锌离子传感器。本发明公开了其制法。
Description
技术领域
本发明涉及1,10-菲罗啉3,8位扩展双-5-甲基噻吩化合物及其在高灵敏锌离子荧光传感与细胞成像等方面的应用。
背景技术
一些重要的阳离子,特别是重金属离子,如Hg2+离子,在自然界可转化为具有神经毒性的甲基汞,通过生物积累造成环境污染和对人畜细胞代谢和神经组织的破坏。再如Cd2+离子,也具有很强的毒性,能够伤害肺,肾脏,脾脏等器官组织;Pb2+离子可导致贫血、肌肉麻痹和智力缺陷;Cu2+离子既可以参与很多生物体内重要的基本生理过程,同时过量的Cu2+离子的存在又会对生物体产生毒性;Zn2+离子既可以参与许多重要的细胞活动,如神经传递、信号传导和基因表达等,但过多的锌离子又会造成农业和食品的污染。所以,对这些离子的重金属的检测显得格外重要。
传统的检测方法,如滴定,原子吸收光谱(AAS),气相或液相色谱等方法,不仅耗时耗力,而且难以做到实时检测乃至于活体检测。因而人们迫切地需要发展出更加简便、准确且能运用于实时、体内和体外的检测方法。为了更方便地监测样品中各种离子浓度,更好地研究这些离子的生理作用,科学家转而开发新型的化学传感器,特别是光化学离子传感器,如紫外-可见光吸收和荧光传感器。这些新型的传感器的显著优点是:灵敏,选择性高,实时,快速响应,成本低,检测限低,无毒无害,甚至可以进行活体内的检测。2000年以后,这方面的研究逐渐成为一个热点,越来越多的金属离子传感器被陆续发明出来,性质越来越好,并逐步向实用化迈进。
在已有的研究与报道中,很大一部分金属离子传感器都是基于有机小分子配体的体系。通过改造原有的一些染料分子或者构建全新的分子骨架,科学家通过在溶液中这些分子对特定金属离子的选择性配位识别,产生明显的光谱变化,继而转化为光信号或电信号,实现化学传感器的作用。一些典型的金属离子传感器如下所示:
1,10-菲罗啉具有大的共轭平面以及强的螯合能力,常被用做双齿螯合配体和金属离子作用形成配合物,受到广泛研究。然而,把1,10-菲罗啉衍生物应用到过渡金属离子传感,并且用于活体细胞成像的案例目前研究的很少,而且多为1:1的金属配合物,具体参见下面两篇相关文献:Y. Li, L. Shi, L. Qin and
Y. Long, Chem. Commun. 2011, 47,
4361-4361.和H. Chen, W. Gao, M. Zhu and Y. Li, Chem. Commun. 2010, 46, 8389-8391.
发明内容
本发明人在已有工作基础上((a) B. Hu,
S. Fu, F. Xu, T. Tao, H. Zhu, K. Cao, W. Huang and X.
You, J. Org. Chem. 2011, 76, 4444-4456; (b) K. Araki, H. Endo, G.
Masuda and T. Ogawa, Chem. Eur. J. 2004, 10, 3331-3340; (c) W.
Huang, H. Tanaka and T. Ogawa, J. Phys. Chem. C 2008, 112,
11513-11526; (d) W. Huang, L. Wang, H. Tanaka and T. Ogawa, Eur. J. Inorg. Chem. 2009, 1321-1330.),设计并合成了一种以1,10-菲罗啉衍生物为基的锌离子荧光传感器,具体技术方案如下:
一种1,10-菲罗啉衍生物,它是3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉(PHT1),具有如下结构式:
一种制备上述3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉的方法,它包括以下步骤:
步骤1. 向100 mL干燥的三颈瓶中加入 0.71 g (29.21 mmol)活化的镁屑、5 mL无水四氢呋喃和催化量的碘,在室温和氩气保护的条件不断的搅拌;将1.7 mL(15.10
mmol)2-溴-5-甲基噻吩溶于5 mL无水四氢呋喃中配成溶液,然后缓慢滴入约10%的2-溴-5-甲基噻吩溶液到上述三颈瓶中,等反应引发后继续滴入剩余的溶液,并保持反应体系处于微沸,滴完后,加热回流30 min;
步骤2. 将步骤1的反应液冷却至室温,倒入50 mL的滴液漏斗中,然后将其缓慢滴入装有2.01 g (5.95 mmol) 3,8-二溴-1,10-菲罗啉、0.09 g(0.17 mmol)
Ni(dppp)Cl2 和 50 mL干燥四氢呋喃的三颈瓶中,滴完后在室温下搅拌2 h,然后加热回流12 h,冷却至室温,加入大量的饱和NH4Cl溶液淬灭,用CHCl3萃取,有机层用饱和食盐水充分洗涤之后用硅胶柱分离(洗脱剂,氯仿:石油醚 = 1:1),旋干溶剂后得到亮黄色的产品3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉。
PHT1为一新化合物,在乙醇-水混合溶液中,PHT1能选择性螯合游离锌离子,形成化学计量比2:1的锌离子配合物([Zn(PHT1)2]2+),配合物稳定常数高达3.4 × 1012 M-1,且在蓝光区域(455 ~ 490 nm)产生51倍的荧光增强。这种荧光增强既不受碱金属离子(如Na+, K+),碱土金属离子(如Ca2+, Mg2+)以及大部分过渡金属离子(如Hg2+, Pb2+,
Ag+)等影响,也不受pH影响,表明PHT1具有高的选择性以及pH稳定性。PHT1对游离锌离子的检测限可以达到5 ppb,远低于世界卫生组织饮用水中的锌含量(3 mg/L)。HL-60细胞、HepG-2细胞的成像实验表明PHT1具有良好的细胞渗透性以及低的细胞毒性,而且能够标记细胞器中痕量的锌离子。上述结果表明PHT1是一种具有高选择性以及高灵敏度的锌离子传感器。
附图说明
图1为化合物PHT1的1H NMR (a), 13C
NMR (b)核磁共振波谱和正离子电喷雾质谱(c)图。
图2为PHT1·C2H5OH化合物的X-射线衍射分析的晶体结构图。
图3为PHT1螯合游离锌离子,形成化学计量比2:1的锌离子配合物([Zn(PHT1)2]2+)示意图及荧光增强照片。
图4为四个1,10-菲罗啉类化合物(10 μM)与1当量Zn2+化合物的荧光激发(a)与发射(b)谱图,其中Phen为1,10-菲罗啉,PHT0为3,8-双-(2-噻吩基)-1,10-菲罗啉,PHT2为3,8-双-(3-甲基-2-噻吩基)1,10-菲罗啉。
图5为PHT1(10 μM)溶液[CH3CH2OH/H2O
(9:1, v/v)]中连续滴加Zn2+离子的荧光发射谱图。
图6为Zn2+ 对于其它金属离子的竞争性实验谱图(Mix1 = Na+, K+,
Li+; Mix2 = Mg2+, Ca2+. Red bars: 10 μM PHT1 + 10 μM metal ions of interest;
Dark bars: 10 μM PHT1 + 10 μM metal ions of interest +
10 μM Zn2+, λex = 405 nm)。
图7为HL-60细胞中不加(左边)入和加入(右边)50 μM Zn2+离子溶液在37 ℃下共同孵育30分钟的激光共聚焦扫描显微镜照片比较。
具体实施方式
实施例
1. 3,8-
双
-
(
5-
甲基
-2-
噻吩基)
-1,10-
菲罗啉的合成
向100 mL干燥的三颈瓶中加入 0.71 g (29.21 mmol)活化的镁屑、5 mL无水四氢呋喃和催化量的碘,在室温和氩气保护的条件不断的搅拌。将1.7 mL(15.10
mmol)2-溴-5-甲基噻吩溶于5 mL无水四氢呋喃中配成溶液,然后缓慢滴入约10% 溶液到上述三颈瓶中,等反应引发后继续滴入剩余的溶液,并保持反应体系处于微沸。滴完后,加热回流30 min。将反应液冷却至室温,倒入50 mL的滴液漏斗中,然后缓慢滴入装有2.01 g (5.95 mmol) 3,8-二溴-1,10-菲罗啉、0.09 g(0.17 mmol)
Ni(dppp)Cl2 和 50 mL干燥四氢呋喃的三颈瓶中,滴完后在室温下搅拌2 h,然后加热回流12 h,冷却至室温,加入大量的饱和NH4Cl溶液淬灭,用CHCl3萃取,有机层用饱和食盐水充分洗涤之后用硅胶柱分离(洗脱剂,氯仿:石油醚
X-射线单晶结构分析表面,噻吩和邻菲咯啉环的二面角很小(图2a),而且分子间的堆积结构显示很强的π-π堆积作用,邻近芳环的质心质心间距在3.6~3.8 Å之间(图2b)。与Phen,PHT0和PHT2的相比:随着推电子噻吩衍生物基团以及推电子甲基基团的引入,综合考虑电子效应和位阻效应,PHT1具有最好的共轭性能和荧光传感性能,具体波谱讨论如下所述。
如图4a所示,四种1,10-菲罗啉化合物的Zn2+配合物的最大激发波长分别为:Phen 385 nm,PHT0 381 nm,PHT1 405 nm,PHT2 375 nm。分别以最大激发波长来进行激发,所得荧光发射光谱如图4b所示,Phen-Zn2+配合物没有荧光,然而PHT0,PHT1以及PHT2的Zn2+配合物分别在440 nm,449 nm,461 nm有强的荧光发射,其中PHT1的荧光量子产率最大。
PHT1乙醇-水溶液的荧光滴定光谱如图5所示,随着Zn2+含量的增加,溶液在461 nm处的荧光发射强度呈线性增强,当Zn2+含量达到0.8当量时,荧光强度达到最大值,通过线性拟合得到方程:I461 nm = 7.41[Zn2+] +
4.83, r = 0.999, n = 11。从图5推算PHT1对溶液中游离Zn2+的检测限为5 ppb,[Zn(PHT1)2]2+配合物的配合常数Kass = 3.4 × 1012 M-2。
不同金属离子的竞争性实验如图6所示,研究对象分别为生命体重常见的碱金属离子(Na+, K+,
Li+),碱土金属离子(Mg2+, Ca2+)以及部分过渡金属离子(Zn2+,Cd2+,Cu2+等)。红色栏表示10 μM PHT1溶液对于1当量不同金属离子的荧光响应,结果表明只有Zn2+和Cd2+导致荧光增强,其中Zn2+的荧光增强最明显。黑色栏表示向已经存在10 μM PHT1以及10 μM Zn2+的溶液中滴加1当量杂离子时荧光响应,结果表明只有Cu2+导致了溶液的荧光猝灭,而其他金属离子没有任何影响。竞争性实验的结果证明了PHT1对Zn2+有良好的选择性。
实施例
2. HL-60
细胞成像
HL-60(人类急性白血病细胞)悬浮液采用DMEM培养基,外加10% FBS(小牛血清)、青霉素(100 μg•mL-1)、链霉素(100 μg•mL-1),于37 ℃和5% CO2气氛下培养。实验中先离心悬浮液并使用Hank's平衡盐溶液(HBSS)清洗,之后用HBSS重新悬浮,与50 μM PHT1在37 ℃下孵育45 min。孵育完毕之后用HBSS洗去剩余PHT1,将其分装在两个培养瓶中,每瓶1 mL细胞悬浮液。其中一瓶加入Zn2+溶液([Zn2+] = 50 μM),之后两瓶悬浮液再在37 ℃下共同孵育20 min。使用激光共聚焦扫描显微镜(TCS SP5, Leica, Germany)分别对两瓶悬浮液进行细胞成像:405 nm激光二极管用于激发光光源,450 nm-500 nm滤光片用于发射光采集。
图7中的两幅图表示的是经过PHT1染色后的HL-60细胞在405 nm激发下的激光共聚焦扫描显微镜照片,微弱的蓝色荧光是由于细胞中含有的痕量Zn2+所导致的。下面两幅图表示的是HL-60细胞经过PHT1染色后,再加入50 μM Zn2+离子溶液孵育30分钟后的激光共聚焦扫描显微镜照片,荧光相对增强表明外加Zn2+透过细胞膜后与细胞内的PHT1结合生成了[Zn(PHT1)2]2+配合物。HL-60细胞的成像实验表明PHT1具有良好的细胞渗透性以及低的细胞毒性,而且能够标记细胞器中痕量的锌离子。
实施例
3. HepG-2
细胞成像
HepG-2(人肝癌细胞)悬浮液采用DMEM培养基,外加10% FBS(小牛血清)、青霉素(100 μg•mL-1)、链霉素(100 μg•mL-1),于37 ℃和5% CO2气氛下培养。染色之前先用DMEM培养基洗两次,再用10 μM PHT1溶液重新悬浮,室温下孵育30 min。孵育完毕之后用DMEM洗去剩余PHT1,将其分装在两个培养瓶中,每瓶1 mL细胞悬浮液。其中一瓶加入Zn2+溶液([Zn2+] = 10 μM),室温下孵育30 min。最后再用锌离子螯合剂(TPEN)处理HepG-2细胞。使用激光共聚焦扫描显微镜进行细胞成像:405 nm激光二极管用于激发光光源,450 nm-500 nm滤光片用于发射光采集。
HepG-2细胞成像的实验结果和结论和HL-60细胞成像的结果类似。
Claims (3)
2. 一种制备权利要求1所述的3,8-双-(5-甲基-2噻吩基)-1,10-菲罗啉的方法,其特征是它包括以下步骤:
步骤1. 向100 mL干燥的三颈瓶中加入 0.71 g
(29.21 mmol)活化的镁屑、5 mL无水四氢呋喃和催化量的碘,在室温和氩气保护的条件不断的搅拌;将1.7
mL(15.10 mmol)2-溴-5-甲基噻吩溶于5 mL无水四氢呋喃中配成溶液,然后缓慢滴入约10%的2-溴-5-甲基噻吩溶液到上述三颈瓶中,等反应引发后继续滴入剩余的溶液,并保持反应体系处于微沸,滴完后,加热回流30 min;
步骤2. 将步骤1的反应液冷却至室温,倒入50 mL的滴液漏斗中,然后将其缓慢滴入装有2.01 g (5.95 mmol)
3,8-二溴-1,10-菲罗啉、0.09 g(0.17
mmol) Ni(dppp)Cl2 和 50 mL干燥四氢呋喃的三颈瓶中,滴完后在室温下搅拌2 h,然后加热回流12 h,冷却至室温,加入大量的饱和NH4Cl溶液淬灭,用CHCl3萃取,有机层用饱和食盐水充分洗涤之后用硅胶柱分离(洗脱剂,氯仿:石油醚 = 1:1),旋干溶剂后得到亮黄色的产品PHT1 。
3. 权利要求1所述的3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉在锌离子传感器中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201110359992.3A CN102603733B (zh) | 2011-12-21 | 2011-12-21 | 3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉及其制法和在锌离子荧光传感与细胞成像中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201110359992.3A CN102603733B (zh) | 2011-12-21 | 2011-12-21 | 3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉及其制法和在锌离子荧光传感与细胞成像中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102603733A true CN102603733A (zh) | 2012-07-25 |
CN102603733B CN102603733B (zh) | 2014-10-01 |
Family
ID=46521563
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201110359992.3A Expired - Fee Related CN102603733B (zh) | 2011-12-21 | 2011-12-21 | 3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉及其制法和在锌离子荧光传感与细胞成像中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102603733B (zh) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104792756A (zh) * | 2015-05-20 | 2015-07-22 | 东南大学 | 四-对-磺酸基-苯基卟啉衍生物作为荧光探针在检测锌离子方面的应用 |
CN106565752A (zh) * | 2016-10-11 | 2017-04-19 | 华南理工大学 | 一种荧光化合物的合成及其在镍离子检测中的应用 |
CN107860755A (zh) * | 2017-11-03 | 2018-03-30 | 苏州科技大学 | 一类邻菲啰啉衍生物、锌离子荧光探针及其应用 |
CN114317684A (zh) * | 2021-12-15 | 2022-04-12 | 南京大学 | 一种基于tna分子的细胞内镁离子成像的方法 |
CN114437118A (zh) * | 2022-01-26 | 2022-05-06 | 南京邮电大学 | 一种噻吩基菲咯啉铕(ⅲ)配合物材料及其制备方法和应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040265628A1 (en) * | 2003-04-17 | 2004-12-30 | Suning Wang | Organic luminescent compounds and methods of making and using same |
WO2009026251A1 (en) * | 2007-08-17 | 2009-02-26 | The General Hospital Corporation | Detecting ions and measuring ion concentrations |
-
2011
- 2011-12-21 CN CN201110359992.3A patent/CN102603733B/zh not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040265628A1 (en) * | 2003-04-17 | 2004-12-30 | Suning Wang | Organic luminescent compounds and methods of making and using same |
WO2009026251A1 (en) * | 2007-08-17 | 2009-02-26 | The General Hospital Corporation | Detecting ions and measuring ion concentrations |
Non-Patent Citations (2)
Title |
---|
KOITI ARAKI ET AL.: "BridgingNanogapElectrodesbyInSituElectropolymerizationofa Bis(terthiophenylphenanthroline)rutheniumComplex", 《CHEM.EUR.J.》, vol. 10, 29 April 2004 (2004-04-29), pages 3331 - 3340 * |
WU ET AL.: "SynthesisandIon-Sensing Phenomena of Two New Helical ConjugatedOligomersContaining1,10-Phenanthroline andOligo-Alkylthiophene", 《JOURNAL OF POLYMER SCIENCE: PART A: POLYMER CHEMISTRY》, vol. 46, 31 December 2008 (2008-12-31), pages 1586 - 1597 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104792756A (zh) * | 2015-05-20 | 2015-07-22 | 东南大学 | 四-对-磺酸基-苯基卟啉衍生物作为荧光探针在检测锌离子方面的应用 |
CN106565752A (zh) * | 2016-10-11 | 2017-04-19 | 华南理工大学 | 一种荧光化合物的合成及其在镍离子检测中的应用 |
CN106565752B (zh) * | 2016-10-11 | 2018-06-19 | 华南理工大学 | 一种荧光化合物的合成及其在镍离子检测中的应用 |
CN107860755A (zh) * | 2017-11-03 | 2018-03-30 | 苏州科技大学 | 一类邻菲啰啉衍生物、锌离子荧光探针及其应用 |
CN107860755B (zh) * | 2017-11-03 | 2020-04-17 | 苏州科技大学 | 一类邻菲啰啉衍生物、锌离子荧光探针及其应用 |
CN114317684A (zh) * | 2021-12-15 | 2022-04-12 | 南京大学 | 一种基于tna分子的细胞内镁离子成像的方法 |
CN114317684B (zh) * | 2021-12-15 | 2023-12-26 | 南京大学 | 一种基于tna分子的细胞内镁离子成像的方法 |
CN114437118A (zh) * | 2022-01-26 | 2022-05-06 | 南京邮电大学 | 一种噻吩基菲咯啉铕(ⅲ)配合物材料及其制备方法和应用 |
Also Published As
Publication number | Publication date |
---|---|
CN102603733B (zh) | 2014-10-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Gu et al. | Synergistic regulation of effective detection for hypochlorite based on a dual-mode probe by employing aggregation induced emission (AIE) and intramolecular charge transfer (ICT) effects | |
Wang et al. | A chemical probe capable for fluorescent and colorimetric detection to Cu2+ and CN− based on coordination and nucleophilic addition mechanism | |
CN102603733B (zh) | 3,8-双-(5-甲基-2-噻吩基)-1,10-菲罗啉及其制法和在锌离子荧光传感与细胞成像中的应用 | |
CN107936262B (zh) | 一种超分子聚合物框架材料的制备及应用 | |
Atilgan et al. | A near IR di-styryl BODIPY-based ratiometric fluorescent chemosensor for Hg (II) | |
Xu et al. | A highly sensitive and selective colorimetric and off–on fluorescent chemosensor for Cu2+ based on rhodamine B derivative | |
Lim et al. | Highly selective fluorescent probe for switch-on Al3+ detection and switch-off F− detection | |
Liu et al. | A novel fluorescent ‘off-on-off’probe for relay recognition of Zn 2+ and Cu 2+ derived from N, N-bis (2-pyridylmethyl) amine | |
Bhatti et al. | New water soluble p-sulphonatocalix [4] arene chemosensor appended with rhodamine for selective detection of Hg2+ ion | |
Park et al. | A new coumarin-based chromogenic chemosensor for the detection of dual analytes Al 3+ and F− | |
Zong et al. | A “turn-on” fluorescence probe towards copper ions based on core-substitued naphthalene diimide | |
Kursunlu et al. | On/off rhodamine-BODIPY-based fluorimetric/colorimetric sensor for detection of mercury (II) in half-aqueous medium | |
Kumar et al. | Highly selective fluorescent probe for detection and visualization of palladium ions in mixed aqueous media | |
Dong et al. | A new colorimetric and fluorescent chemosensor based on Schiff base-phenyl-crown ether for selective detection of Al3+ and Fe3+ | |
Li et al. | A rhodamine-benzimidazole based chemosensor for Fe 3+ and its application in living cells | |
Na et al. | Benzothiazole-based heterodipodal chemosensor for Cu2+ and CN–ions in aqueous media | |
Yan et al. | A new dual-function fluorescent probe of Fe3+ for bioimaging and probe-Fe3+ complex for selective detection of CN− | |
CN106957308B (zh) | 基于香豆素类的接力响应型荧光分子探针及其制备方法和用途 | |
Piyanuch et al. | Rapid and visual detection of Cd2+ based on aza-BODIPY near infrared dye and its application in real and biological samples for environmental contamination screening | |
Wang et al. | New fluorescent chemosensors based on mononuclear copper complex for highly selective and sensitive detection of phosphate anion in aqueous solution and living cells | |
Yang et al. | Efficient energy transfer in a tri-chromophoric dyad containing BODIPYs and corrole based on a truxene platform | |
Li et al. | Highly selective and reversible colorimetric detection of mercury ions by a hydrophilic cycloruthenated complex in water | |
Zhao et al. | A FRET-based ratiometric fluorescent probe for Hg2+ detection in aqueous solution and bioimaging in multiple samples | |
Biswal et al. | A pyridine and pyrrole coupled rhodamine derivative for Co (II) ion detection and its imaging application in plant tissues | |
Dumanoğulları et al. | Investigation of ultrafast energy transfer mechanism in BODIPY–Porphyrin dyad system |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141001 Termination date: 20141221 |
|
EXPY | Termination of patent right or utility model |