A fluorescent probe for detecting butyrylcholinesterase activity and its preparation method and application
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A butyrylcholinesterase and fluorescent probe technology, applied in the field of fluorescent probes, can solve problems such as no public detection function
Active Publication Date: 2022-06-10
ZHEJIANG NORMAL UNIVERSITY
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[0004] Patent No. 2013106811585 discloses a long-wavelength fluorescent probe and its preparation method and application. The compound used as the probe is 1,7-dimethyl-3,5-distyryl-8-benzene Base-(2'-maleimide)-boron difluoride dipyrromethane has good selectivity to sulfhydryl compounds, and the fluorescence intensity increases after reacting with sulfhydryl groups, but the long-wavelength fluorescent probe is not disclosed in the patent. BChE has detection function
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Embodiment 1
[0044] Two, 2,3-dicyano-1,4-phenylene diacrylate (DCPDA) to the fluorescence response of sulfhydryl compounds.
[0055] DCPDA, butyrylthiocholine and HEPES buffer. Apply the above fluorescence detection kit to butyrylcholinester
[0058] Except adding GSH at a concentration of 10.0 μM, 20.0 μM, 30.0 μM or 100.0 μM before adding 1.0 mM BTh,
[0061] In order to verify the function of BChE detection inhibitor screening, take tacrine as an example. Inhibition of BChE activity by tacrine
Embodiment 2
[0078] DCPDMA, butyrylthiocholine and HEPES buffer. Apply the above fluorescence detection kit to butyrylcholinester
[0079] First prepare a stock solution of DCPDMA solution in THF, wherein the DCPDMA solution is 10.0 mM; then mix 30.0 L
[0081] In addition to adding an amount of GSH (10.0 μM, 20.0 μM, 30.0 μM or 100.0 μM) prior to the addition of BTCh (1.0 mM)
[0082] The selective detection of BChE from mixtures with AChE was carried out as follows: containing specific levels of different species
[0083] In order to verify the function of BChE detection inhibitor screening, take tacrine as an example. Inhibition of BChE activity by tacrine
Embodiment 3
[0090] To 3.0 [mu]L of HEPES buffer was added 90 [mu]L of a 10.0 mM solution of DCPDFC. dissolved in the above DCPDFC
[0091] First prepare a stock solution of DCPDFC solution in THF, wherein the DCPDFC solution is 10.0 mM; then mix 30.0 μL
[0092] The experiment of the effect of GSH on the BChE assay was as follows: except that a certain
[0095] In order to verify the function of BChE detection inhibitor screening, take tacrine as an example. Inhibition of BChE activity by tacrine
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Abstract
The invention discloses a fluorescent probe for detecting the activity of butyrylcholinesterase, the molecular structure is as follows: the preparation method of the fluorescent probe comprises the following steps: step 1, dissolving 2,3-dicyanohydroquinone in In dry tetrahydrofuran; step 2, dropwise adding excessive acryloyl chloride or 3,3-dimethylacryloyl chloride or a kind of 4,4,4-trifluorocrotonoyl chloride to the mixed solution of step 1; step 3, adding Add a small amount of triethylamine dropwise to the mixed solution in step two; step four, stir the mixed solution in step three; remove the solvent with a rotary evaporator, purify the product with silica gel column chromatography, and finally obtain a white powder; The needle is used to detect the activity of butyrylcholinesterase; the invention can be illuminated by thiol through a fast mercaptoene click reaction to achieve fluorescence enhancement, with good selectivity and strong anti-interference.
Description
Fluorescent probe for detecting butyrylcholinesterase activity and preparation method and application thereof technical field The present invention relates to fluorescent probe technical field, particularly relate to a kind of fluorescent probe that detects butyrylcholinesterase activity Needles and methods of making and using the same. Background technique Thiol-reactive probes are commonly used to label proteins to monitor conformational changes, assembly of multiple subunits, and ligand binding process, and is often used to detect small biothiols such as glutathione, which are important in human health and disease play an important special role. At present, the existing thiol-reactive fluorescent probes have the disadvantage that they are not suitable for all thiol-containing substances. No specific selectivity, there is an urgent need to design and develop new selective recognition fluorophores for glutathione, cysteine and homocysteine. light probe. Among ...
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