Near infrared fluorescent probe for detecting zinc ion in cell and its synthesis method and use

Abstract

The present invention provides the near-infrared fluorescent probe that detects intracellular zinc ion and synthetic method and purposes, and this synthetic method is carried out as follows: a. with the weight of cyanine dye Cy.7: complexing group=1: 5~10 The ratio of parts is to add N,N-dimethylformamide to dissolve the raw materials, and then react for 2-5 hours under the protection of an inert gas at 60-70°C; b. Cool the reaction mixture obtained in step a to room temperature to 10-20°C , evaporated to dryness under reduced pressure, then dissolved with diethyl ether, then added water to extract until layered, took the extract I and added ethyl acetate to extract until layered, and obtained extract II; c. Take the extract II obtained in step b and add anhydrous sulfuric acid Magnesium is dried, and then evaporated to dryness under reduced pressure at 30-40°C, and ethyl acetate is evaporated to obtain the product. The near-infrared fluorescent probe has good cell permeability, has no toxic and side effects on the cells themselves, and is particularly suitable for the detection of intracellular zinc ions.

Description

technical field [0001] The present invention relates to the fields of biological detection technology and clinical medical detection, in particular to a near-infrared fluorescent probe for detecting intracellular zinc ions; the present invention also relates to a synthesis method of the fluorescent probe; in addition, the present invention also relates to the fluorescent probe the use of. Background technique [0002] Zinc ions are the second most abundant transition metal in living organisms. A large number of zinc ions are concentrated in nerve tissue, such as the concentration in brain tissue is 0.1-0.5nM. Zinc ions are directly involved in the regulation of gene expression through metal regulatory proteins. For example, zinc, as a component of zinc finger proteins, can affect gene expression by changing the structure of chromatin or by interrupting DNA synthesis. Most of the zinc ions in organisms are tightly combined with proteins, and there are "free zinc pools" in s...

AI Technical Summary

Problems solved by technology

[0003] Many existing methods, including atomic absorption spectrometry, electron paramagnetic resonance, Electron Probe Microanalysis, etc., have been used for the detection of metal ions in cells, but the instruments used in these methods are expensive, and they cannot perform real-time and original detection of metal ions in organisms. Bit dynamic detection, and the sample pretreatment is more complicated when these methods are determined, so the application is limited

[0004] At present, existing fluorescent probes that can detect cellular zinc ions mainly include the following categories: TSQ (Frederickson, C.J.; Kasarkis, E.J.; Ringo, D.; Frederickson, R.E.J.Neurosci.Methods, 1987,20,91- 103.), Zinquin ethyl ester (Tsuda, M.Neurosci., 17, 6678, 1997) dansylaminoethyl cyclamate (Koike, T.; Watanabe, T.; Aoki, S.; Kimura, E. ; Shiro, M., J.Am.Chem.Sic., 118, 12696-12703, 1996) zinpyr (Burfrtte, S.C.; Frederickson, C.J.; Bu, W.; Lippard, S.J., J.Am.Chem.Soc. , 2003, 125, 1778-1787.) and ZnAF (Hirano, T.; Kikuchi, K.; Urano, Y.; Higuchi, T.; Nagano, T.J.Am.Chem.Soc., 2002, 124, 6555-6562 .) etc., but the disadvantage of these probes is that their excitation wavelength is in the ultraviolet region (~350nm), which will cause damage to the cells when excited, and because their emission wavelength is in the ultraviolet or visible region, the autofluorescence interference of the biological body is serious, and the pH change Sensitive, making it subject to certain restrictions

Another problem is that it is generally difficult to synthesize such probe molecules, and the structure is relatively complex, so it will be difficult for practical application.

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