CN102603733A - 3, 8-bis-(5-methyl-2-thienyl)-1, 10-phenanthroline, preparation method thereof and application in zinc ion fluorescent sensing and cell imaging - Google Patents
3, 8-bis-(5-methyl-2-thienyl)-1, 10-phenanthroline, preparation method thereof and application in zinc ion fluorescent sensing and cell imaging Download PDFInfo
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- 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
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- 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
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- 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
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- 238000009835 boiling Methods 0.000 claims description 3
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- 150000005045 1,10-phenanthrolines Chemical class 0.000 abstract description 2
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- YNPNZTXNASCQKK-UHFFFAOYSA-N Phenanthrene Natural products C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 3
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Images
Abstract
The invention relates to a 1, 10-phenanthroline derivative, in particular to a 3, 8-bis-(5-methyl-2-thienyl)-1, 10-phenanthroline (PHT1), which has a structural formula as shown in the accompanying drawing. In ethanol-water mixed solution, the PHT1 can be selectively chelated with free zinc ions to form zinc ion complex ([Zn(PHT1)2]2+) with stoichiometric ratio being 2:1. The stability constant of the complex highly reaches 3.4*1012M<-1> and 51 times of fluorescence enhancement can be generated in a blue light area (455-490nm). The fluorescence enhancement not only is not influenced by alkali metal ions such as Na+ and K+, alkaline-earth metal ions such as Ca2+ and Mg2+, most transition metal ions such as Hg2+, Pb2+ and Ag+ and the like, but also is not influenced by pH, and the detection limit to free zinc ions can reach 9ppb. According to imaging experiments of HL-60 cells and HepG-2 cells, the PHT1 has good cell permeability and low cell toxicity, can label trace zinc ions in cell organs and can be used in zinc ion sensors. The invention additionally discloses a preparation method of the PHT1.
Description
Technical field
The present invention relates to 1, the expansion of 3,8 of 10-phenanthrolines is two-5-thiotolene compound and in the application of aspects such as highly sensitive zine ion fluorescence sense and cell imaging.
Background technology
The positively charged ion that some are important, particularly heavy metal ion are like Hg
2+Ion can be converted at nature and to have neurovirulent Methylquecksilber, causes environmental pollution and to the destruction of people and animals' cellular metabolism and nervous tissue through biological accumulation.Cd for another example
2+Ion also has very strong toxicity, can injure organ-tissues such as lung, kidney, spleen; Pb
2+Ion can cause anaemia, muscular paralysis and amentia; Cu
2+Ion both can have been participated in basic physiological process important in a lot of organisms, simultaneously excessive Cu
2+Ionic exists again can be to the organism toxigenicity; Zn
2+Ion both can have been participated in many important cellular activities, and like neurotransmission, signal conduction and genetic expression etc., but too much zine ion can cause the pollution of agricultural and food again.So, to the detection of these ionic heavy metals important that seems.
Traditional detection method, like titration, atomic absorption spectrum (AAS), methods such as gas phase or liquid chromatography not only take time and effort, and even and are difficult to accomplish detect in real time live body and detect.Thereby people need develop urgently easier, accurately and can apply in real time, in the body and external detection method.In order to monitor various ionic concns in the sample more easily, study these ionic physiological actions better, scientist transfers the chemical sensor, particularly photochemistry ionization sensor of development of new, like ultraviolet-visible photoabsorption and fluorescent optical sensor.The remarkable advantage of the transmitter that these are novel is: sensitivity, and selectivity is high, in real time, and response fast, cost is low, and detectability is low, and is nontoxic, even can carry out intravital detection.After 2000, the research of this respect becomes a focus gradually, and increasing metal ion sensor is invented successively, and character is become better and better, and progressively strides forward to practicability.
In existing research and report, metal ion sensor all is based on the system of organic molecule part greatly.Through transforming more original dye molecules or making up brand-new molecular skeleton; Scientist discerns special metal ionic selective coordination through these molecules in solution; Produce tangible spectrum change, be converted into optical signal or electrical signal then, realize the effect of chemical sensor.Some typical metal ion sensors are as follows:
1, the 10-phenanthroline has big conjugate planes and strong sequestering power, often is used as double-tooth chelate ligand and metals ion effect and forms title complex, receives broad research.Yet 1,10-phenanthroline verivate is applied to the transition metal ion sensing; And the case that is used for the active somatic cell imaging is studied seldom at present, and is mostly the metal complexes of 1:1, specifically referring to following two pieces of pertinent literatures: Y. Li; L. Shi, L. Qin and Y. Long
Chem. Commun.2011,
47, 4361-4361. and H. Chen, W. Gao, M. Zhu and Y. Li,
Chem. Commun.2010,
46, 8389-8391.
Summary of the invention
The inventor on existing working foundation ((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. C2008,
112, 11513-11526; (d) W. Huang, L. Wang, H. Tanaka and T. Ogawa,
Eur. J. Inorg. Chem. 2009,1321-1330.), designed and synthesized a kind ofly with 1,10-phenanthroline verivate is the zine ion fluorescent optical sensor of base, concrete technical scheme is following:
A kind of 1,10-phenanthroline verivate, it is 3,8-is two-(5-methyl-2-thienyl)-1,10-phenanthroline (PHT1) has following structural formula:
A kind of preparation is above-mentioned 3, and 8-is two-(5-methyl-2-thienyl)-1, the method for 10-phenanthroline, it may further comprise the steps:
Step 1. adds the iodine of 0.71 g (29.21 mmol) activatory magnesium chips, 5 mL anhydrous tetrahydro furans and catalytic amount in 100 mL exsiccant three-necked bottles, constantly stir in the condition of room temperature and argon shield; 1.7 mL (15.10 mmol) 2-bromo-5-thiotolene is dissolved in wiring solution-forming in the 5 mL anhydrous tetrahydro furans; Slowly splash into about 10% 2-bromo-5-thiotolene solution then in above-mentioned three-necked bottle; Splash into rest solution Deng reaction initiation continued; And keep reaction system to be in little boiling, and after dripping off, reflux 30 min;
PHT1 is a new compound, in the alcohol-water mixing solutions, and PHT1 ability selectivity chelating free zinc ion, zine ion the title complex ([Zn (PHT1) of formation stoichiometric ratio 2:1
2]
2+), the title complex stability constant is up to 3.4 * 10
12M
-1, and produce 51 times fluorescence in blue region (455～490 nm) and strengthen.This fluorescence enhancing neither receives alkalimetal ion (like Na
+, K
+), alkaline earth metal ion is (like Ca
2+, Mg
2+) and most of transition metal ion (like Hg
2+, Pb
2+, Ag
+) wait influence, not influenced by pH, show that PHT1 has high selectivity and pH stability.PHT1 can reach 5 ppb to the detectability of free zinc ion, far below the zinc content in World Health Organization's tap water (3 mg/L).The imaging experiment of HL-60 cell, HepG-2 cell shows that PHT1 has good cell perviousness and low cytotoxicity, and the zine ion of trace in can the labeled cell device.The above results shows that PHT1 is a kind of have highly selective and highly sensitive zine ion transmitter.
Description of drawings
Fig. 1 is a compound
PHT1 1H NMR
(a),
13C NMR
(b)NMR spectrum and positive ion electrospray ionization mass spectrum (
c) figure.
Fig. 2 does
PHT1C
2H
5The crystalline structure figure that the X-ray diffraction of OH compound is analyzed.
Fig. 3 is a PHT1 chelating free zinc ion, forms zine ion the title complex ([Zn (PHT1) of stoichiometric ratio 2:1
2]
2+) synoptic diagram and fluorescence enhancing photo.
Fig. 4 is four 1,10-phenanthroline compounds (10 μ M) and 1 equivalent Zn
2+The fluorescence excitation of compound (
a) and emission (
b) spectrogram, wherein Phen is 1, and 10-phenanthroline, PHT0 are 3, and 8-pair-(2-thienyl)-1,10-phenanthroline, PHT2 are 3,8-pair-(3-methyl-2-thienyl) 1, the 10-phenanthroline.
Fig. 5 does
PHT1(10 μ M) solution [CH
3CH
2OH/H
2O (9:1,
v/
v)] in drip continuously Zn
2+Ionic fluorescent emission spectrogram.
Fig. 6 is Zn
2+Competitiveness experiment spectrogram (Mix1=Na for other metals ion
+, K
+, Li
+; Mix2=Mg
2+, Ca
2+. Red bars:10 μ M
PHT1+ 10 μ M metal ions of interest; Dark bars:10 μ M
PHT1+ 10 μ M metal ions of interest+10 μ M Zn
2+, λ
Ex=405 nm).
Fig. 7 is not for adding (left side) in the HL-60 cell and go into and add (the right) 50 μ M Zn
2+The laser confocal scanning microscope photo that solion was hatched under 37 ℃ 30 minutes jointly compares.
Embodiment
Embodiment 1. 3, and 8-is two-(5-methyl-2-thienyl)-1,10-phenanthroline synthetic
The iodine that in 100 mL exsiccant three-necked bottles, adds 0.71 g (29.21 mmol) activatory magnesium chips, 5 mL anhydrous tetrahydro furans and catalytic amount constantly stirs in the condition of room temperature and argon shield.1.7 mL (15.10 mmol) 2-bromo-5-thiotolene is dissolved in wiring solution-forming in the 5 mL anhydrous tetrahydro furans, slowly splashes into about 10% solution then in above-mentioned three-necked bottle, wait reaction to cause continued and splash into rest solution, and keep reaction system to be in little boiling.After dripping off, reflux 30 min.Reaction solution is cooled to room temperature, pours in the tap funnel of 50 mL, slowly splash into then 2.01 g (5.95 mmol) 3 are housed, 8-two bromo-1,10-phenanthroline, 0.09 g (0.17 mmol) Ni (dppp) Cl
2In the three-necked bottle of 50 mL dry tetrahydrofuran, at room temperature stir 2 h after dripping off, reflux 12 h are cooled to room temperature then, add a large amount of saturated NH
4CHCl is used in the cancellation of Cl solution
3Extraction, organic layer is with separating (eluent, chloroform: sherwood oil with silicagel column after the saturated aqueous common salt thorough washing
X-ray crystal structure is analyzed the surface, and the interfacial angle of thiophene and o-phenanthroline ring is very little, and (Fig. 2 a), and intermolecular packed structures shows very strong pi-pi accumulation effect, is close to the barycenter barycenter spacing (Fig. 2 b) between 3.6～3.8 of aromatic ring.With Phen; PHT0 and PHT2 compare: along with the introducing that pushes away electronics thiophene derivant group and push away the electronics methyl group; Take all factors into consideration electronic effect and steric effect, PHT1 has best conjugation performance and fluorescence sense performance, and concrete wave spectrum discussion is described below.
Shown in Fig. 4 a, four kind 1, the Zn of 10-phenanthroline compound
2+The maximum excitation wavelength of title complex is respectively: Phen 385 nm, PHT0 381 nm, PHT1 405 nm, PHT2 375 nm.Excite with the maximum excitation wavelength respectively, the gained fluorescence emission spectrum shown in Fig. 4 b, Phen-Zn
2+Title complex does not have fluorescence, however PHT0, the Zn of PHT1 and PHT2
2+Title complex is respectively at 440 nm, 449 nm, and 461 nm have strong fluorescent emission, and wherein the fluorescence quantum yield of PHT1 is maximum.
The fluorometric titration spectrum of PHT1 ethanol-water solution is as shown in Figure 5, along with Zn
2+The increase of content, the fluorescent emission intensity of solution at 461 nm places are linear and strengthen, and work as Zn
2+When content reached 0.8 equivalent, fluorescence intensity reached peak, obtained equation through linear fit:
I 461 nm =7.41 [Zn
2+]+4.83, r=0.999, n=11.Calculate that from Fig. 5 PHT1 is to free Zn the solution
2+Detection be limited to 5 ppb, [Zn (PHT1)
2]
2+The cooperation constant of title complex
K Ass =3.4 * 10
12M
-2
The competitive experiment of different metal ionic is as shown in Figure 6, and research object is respectively the common alkalimetal ion (Na of life body weight
+, K
+, Li
+), alkaline earth metal ion (Mg
2+, Ca
2+) and part transition metal ion (Zn
2+, Cd
2+, Cu
2+Deng).10 μ M PHT1 solution are represented for 1 equivalent different metal ionic fluorescence response in red hurdle, and the result shows to have only Zn
2+And Cd
2+Cause fluorescence to strengthen, wherein Zn
2+Fluorescence strengthen the most obvious.10 μ M PHT1 and 10 μ M Zn are represented to exist in the black hurdle
2+Solution in fluorescence response when dripping 1 equivalent hetero-ion, the result shows to have only Cu
2+The quenching of fluorescence that has caused solution, and other metals ions have no influence.Competitive result of experiment has proved that PHT1 is to Zn
2+Good selectivity is arranged.
HL-60 (human acute leukemia cells) suspension-s adopts the DMEM substratum, adds 10% FBS (calf serum), penicillium mould (100 μ gmL
-1), Streptomycin sulphate (100 μ gmL
-1), in 37 ℃ and 5% CO
2Cultivate under the atmosphere.First centrifugal suspension-s and use Hank's balanced salt solution (HBSS) to clean in the experiment suspends with HBSS afterwards again, under 37 ℃, hatches 45 min with 50 μ M PHT1.Hatching finishes remains PHT1 with the HBSS flush away afterwards, it is divided in two culturing bottles every bottle 1 mL cell suspending liquid.Wherein one bottle adds Zn
2+Solution ([Zn
2+]=50 μ M), two bottles of suspension-s are hatched 20 min more jointly under 37 ℃ afterwards.(TCS SP5, Leica Germany) carry out cell imaging to two bottles of suspension-s respectively: 405 nm laser diodes are used for exciting light sources, and 450 nm-500 nm spectral filters are used to launch light collection to use laser confocal scanning microscope.
What two width of cloth figure among Fig. 7 represented is through the laser confocal scanning microscope photo of HL-60 cell under 405 nm excite after the PHT1 dyeing, and faint blue-fluorescence is because the trace Zn that contains in the cell
2+Caused.Below two width of cloth figure represent is the HL-60 cell through after the PHT1 dyeing, add 50 μ M Zn again
2+Solion is hatched the laser confocal scanning microscope photo after 30 minutes, and fluorescence relatively strengthens and shows and combine to have generated [Zn (PHT1) with intracellular PHT1 after adding the Zn2+ permeate through cell membranes
2]
2+Title complex.The imaging experiment of HL-60 cell shows that PHT1 has good cell perviousness and low cytotoxicity, and the zine ion of trace in can the labeled cell device.
HepG-2 (human liver cancer cell) suspension-s adopts the DMEM substratum, adds 10% FBS (calf serum), penicillium mould (100 μ gmL
-1), Streptomycin sulphate (100 μ gmL
-1), in 37 ℃ and 5% CO
2Cultivate under the atmosphere.Earlier wash twice before the dyeing, suspend again with 10 μ M PHT1 solution again, hatch 30 min under the room temperature with the DMEM substratum.Hatching finishes remains PHT1 with the DMEM flush away afterwards, it is divided in two culturing bottles every bottle 1 mL cell suspending liquid.Wherein one bottle adds Zn
2+Solution ([Zn
2+]=10 μ M), hatch 30 min under the room temperature.Use zine ion sequestrant (TPEN) to handle the HepG-2 cell at last again.Use laser confocal scanning microscope to carry out cell imaging: 405 nm laser diodes are used for exciting light sources, and 450 nm-500 nm spectral filters are used to launch light collection.
The result of the experimental result of HepG-2 cell imaging and conclusion and HL-60 cell imaging is similar.
Claims (3)
1. one kind 1,10-phenanthroline verivate is characterized in that: it is 3, and 8-pair-(5-methyl-2-thienyl)-1, the 10-phenanthroline has following structural formula:
One kind to prepare claim 1 described 3,8-is two-(5-methyl-2 thienyl)-1, the method for 10-phenanthroline is characterized in that it may further comprise the steps:
Step 1. adds the iodine of 0.71 g (29.21 mmol) activatory magnesium chips, 5 mL anhydrous tetrahydro furans and catalytic amount in 100 mL exsiccant three-necked bottles, constantly stir in the condition of room temperature and argon shield; 1.7 mL (15.10 mmol) 2-bromo-5-thiotolene is dissolved in wiring solution-forming in the 5 mL anhydrous tetrahydro furans; Slowly splash into about 10% 2-bromo-5-thiotolene solution then in above-mentioned three-necked bottle; Splash into rest solution Deng reaction initiation continued; And keep reaction system to be in little boiling, and after dripping off, reflux 30 min;
Step 2. is cooled to room temperature with the reaction solution of step 1, pours in the tap funnel of 50 mL, then it is slowly splashed into 2.01 g (5.95 mmol) 3 are housed, 8-two bromo-1,10-phenanthroline, 0.09 g (0.17 mmol) Ni (dppp) Cl
2In the three-necked bottle of 50 mL dry tetrahydrofuran, at room temperature stir 2 h after dripping off, reflux 12 h are cooled to room temperature then, add a large amount of saturated NH
4CHCl is used in the cancellation of Cl solution
3Extraction, organic layer is with (eluent, chloroform: sherwood oil=1:1) obtains jonquilleous product after revolving dried solvent with the silicagel column separation after the saturated aqueous common salt thorough washing
PHT1
3. claim 1 is described 3, and 8-is two-(5-methyl-2-thienyl)-1, the application of 10-phenanthroline in the zine ion transmitter.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792756A (en) * | 2015-05-20 | 2015-07-22 | 东南大学 | Application of tetra-p-sulfonic group-phenyl porphyrin derivative as fluorescent probe in aspect of detecting zinc ions |
| CN106565752A (en) * | 2016-10-11 | 2017-04-19 | 华南理工大学 | Synthesis of fluorescent compound and applications of fluorescent compound in nickel ion detection |
| CN107860755A (en) * | 2017-11-03 | 2018-03-30 | 苏州科技大学 | Phenanthroline derivative, zinc ion fluorescent probe and application thereof |
| CN114317684A (en) * | 2021-12-15 | 2022-04-12 | 南京大学 | Intracellular magnesium ion imaging method based on TNA molecules |
| CN114437118A (en) * | 2022-01-26 | 2022-05-06 | 南京邮电大学 | Thienyl phenanthroline europium (III) complex material and preparation method and application thereof |
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/en 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 |
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| CN104792756A (en) * | 2015-05-20 | 2015-07-22 | 东南大学 | Application of tetra-p-sulfonic group-phenyl porphyrin derivative as fluorescent probe in aspect of detecting zinc ions |
| CN106565752A (en) * | 2016-10-11 | 2017-04-19 | 华南理工大学 | Synthesis of fluorescent compound and applications of fluorescent compound in nickel ion detection |
| CN106565752B (en) * | 2016-10-11 | 2018-06-19 | 华南理工大学 | A kind of synthesis of fluorescent chemicals and its application in nickel ion detection |
| CN107860755A (en) * | 2017-11-03 | 2018-03-30 | 苏州科技大学 | Phenanthroline derivative, zinc ion fluorescent probe and application thereof |
| CN107860755B (en) * | 2017-11-03 | 2020-04-17 | 苏州科技大学 | Phenanthroline derivative, zinc ion fluorescent probe and application thereof |
| CN114317684A (en) * | 2021-12-15 | 2022-04-12 | 南京大学 | Intracellular magnesium ion imaging method based on TNA molecules |
| CN114317684B (en) * | 2021-12-15 | 2023-12-26 | 南京大学 | Intracellular magnesium ion imaging method based on TNA molecules |
| CN114437118A (en) * | 2022-01-26 | 2022-05-06 | 南京邮电大学 | Thienyl phenanthroline europium (III) complex material and preparation method and application thereof |
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