Fluoroboric dye fluorescent probe for cell zinc ion detection

A technology of fluorescent probes and zinc ions, which is applied in the direction of luminescent materials, biological testing, material inspection products, etc., can solve the problems of difficult synthesis of probe molecules, complex structures, difficulties, etc., and achieve high yield, no toxic side effects, and stable good sex effect

Inactive Publication Date: 2006-01-04
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of probe is limited in the study of zinc ion changes in environments with small pH changes or large pH changes
Another problem is that it is generally difficult to synthesize such probe molecules, the structure is relatively complex, and large-scale industrialization will be difficult.

Method used

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  • Fluoroboric dye fluorescent probe for cell zinc ion detection
  • Fluoroboric dye fluorescent probe for cell zinc ion detection
  • Fluoroboric dye fluorescent probe for cell zinc ion detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Synthesis of Probe 3

[0032]

[0033] (1) Synthesis of Intermediate 2

[0034] In a 1000ml three-necked flask, add 2.17 g (19.4 mmol) of chloroacetyl chloride and 3.7 g of 1,4-dimethylpyrrole to dissolve in 2 L of dichloromethane, stir at room temperature for 5 hours under nitrogen protection, and evaporate the mixed solution to dryness under reduced pressure. 0.5L, then 40ml of triethylamine was added, and then 80ml of boron trifluoride was added, and the mixture was continuously stirred for 4 hours. The mixture was washed with water, dried and evaporated to dryness. Silica gel column separation (16:1 n-hexane:ethyl acetate) afforded 1.69 g of Intermediate 2. Yield 34%. 1 H-NMR (400MHz, CDCl 3 , ppm): δ2.53 (s, 12H); 4.78 (s, 2H); 6.08 (s, 2H). 13 C-NMR (400MHz, CDCl 3 , ppm): δ14.86, 15.69, 37.33, 122.46, 131.57, 136.14, 141.31, 156.83. 19 F(400MHz, CDCl 3 , CFCl 3 asexternal reference, δ=0): δ-146.73(q, J=36Hz). Anal.Calcd for C 14 H 16 BClF 2 N 2 :...

Embodiment 2

[0038] Probe 3 zinc ion selectivity

[0039] The selectivity to zinc ions was evaluated using compound 3 synthesized above. 1 μM of compound 3 was added to a five-fold excess of various metal ions in Tris-HCl buffer at pH 7.4 (20 mM). The excitation wavelength of probe 3 was 492 nm and the emission wavelength was 511 nm. The test results are shown in In Figure 1 . In the figure, the fluorescence intensity on the vertical axis is set to 1 when the metal ion is not added, and the fluorescence intensity when various metal ions are added is represented by numerical values. It can be seen from the figure that compound 3 has high sensitivity to zinc ions, the addition of zinc ions produces a great fluorescence enhancement, and also has good selectivity to zinc ions, such as sodium, potassium, calcium, magnesium, and manganese. , iron and other metal ions do not interfere with the detection.

Embodiment 3

[0041] Insensitivity of probe 3 to pH

[0042] Use the compound 3 synthesized above to evaluate the response to pH. For compound 3, adjust the pH value to about 1.0 in an aqueous solution with an ionic strength of 0.1. After measuring the fluorescence intensity, add lye to slowly increase the pH value and record the corresponding fluorescence. Intensity changes, test results are shown in In Figure 3 . It can be seen from the figure that the probe 3 has a very low pKa value, and in the pH range of 4 to 10, the pH change has little effect on the fluorescence emission. In addition, the same method was used to measure the pH response of 1:1 zinc ion complex with compound 3. The test results are shown in Figure 3 in the illustration. It can be seen from the figure that in the range of pH 3 to 10, the pH change has little effect on the fluorescence emission. Therefore, probe 3 can be used for the detection of intracellular zinc ions in a wide pH range.

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Abstract

The fluoroboric dye fluorescent probe for cell zinc ion detection operates on inner photoinduced electron transfer (PET) principle. The probe is prepared through connecting fluoroboric dye with 8-site chloromethyl group and bis(2-pyridyl methyl) amine simply. It has exciting wavelength and emitting wavelength in the visible light region, and has excellent chemical and optical stability. The probe series has very low pKa value of 2-3 normally, and has no effect on fluorescent emitting of pH in the range of 4-10. It has high selectivity on zinc ion without interference from Na, K, Ca, Mg, Mn and other metal ions, and can detect zinc ion concentration as low as nanomole level. The present invention has high quantum yield of probe molecule-zinc ion complex. Fluorescent microscopic imaging shows that the probe has high cell permeability and no toxic side effect on cell and is especially suitable for detection of intracellular zinc ion concentration.

Description

technical field [0001] The invention belongs to a fluorescent molecular probe for cell zinc ion detection. Background technique [0002] Zinc ion is a divalent metal ion in organisms, and it is also the second enriched transition metal after relaying iron in organisms. A large amount of zinc ions are concentrated in nerve tissue, such as the concentration in brain tissue is 0.1-0.5mM. Although most zinc ions in organisms are tightly bound to proteins, there are still "free zinc pools" in some cells, and the concentration of free zinc is as low as nanomolar in various mammalian cells. To date, it has been recognized that zinc plays many different roles in life. One of the most important and well-known roles is as a structural factor in metalloproteins. At the same time, zinc ions are directly involved in regulating gene expression through metal regulatory proteins. There is also a lot of evidence in vivo and in vitro that zinc is an important regulatory element in apoptos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52C09K11/00
Inventor 彭孝军吴云扣樊江莉王静云郭斌臣崔爱军田茂忠
Owner DALIAN UNIV OF TECH
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