Enzyme-responsive self-aggregation luminous molecule and applications thereof in monitoring enzyme activity

A self-aggregation luminescent, enzyme-responsive technology, applied in the direction of chemiluminescence/bioluminescence, and analysis by making materials undergo chemical reactions, to achieve excellent biocompatibility, strong biocompatibility, and avoid aggregation-induced luminescence quenching and the effect of photoquenching

Active Publication Date: 2014-01-22
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technical method, which can be applied to both single-cell sites and enzymatic reaction process spatiotemporal monitoring, has not been reported so far.

Method used

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  • Enzyme-responsive self-aggregation luminous molecule and applications thereof in monitoring enzyme activity
  • Enzyme-responsive self-aggregation luminous molecule and applications thereof in monitoring enzyme activity
  • Enzyme-responsive self-aggregation luminous molecule and applications thereof in monitoring enzyme activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] First, this example describes the synthesis of tetraphenylethylene carboxylic acid derivatives, a self-aggregating luminescent molecule, with the following route:

[0039]

[0040] (I) Preparation of I, I, 2-triphenyl-2-Gv-methoxyphenyl)ethene: 3.68g, 20mmol of benzophenone was added to a clean three-neck round bottom flask, and then 19.3g, 100mmol of zinc powder was added. The flask was evacuated and filled with dry nitrogen three times. Afterwards, inject 200mL of anhydrous tetrahydrofuran into the flask, add 6.8mL, 60mmol of titanium tetrachloride dropwise at 0°C, reflux overnight and add 10% potassium carbonate solution (13.8g of potassium carbonate dissolved in 125mL of water) to terminate the reaction. After the phases were separated, the organic phase was washed twice with brine and dried over anhydrous magnesium sulfate, then filtered and concentrated. The obtained crude product was purified by silica gel column, and was eluted with petroleum ether and dieth...

Embodiment 2

[0048] First, synthesize TPE with the same method and process as in Example 1.

[0049] Secondly, in this embodiment, the live cell imaging of protein tyrosine phosphatase 1B (Protein Tyrosine Phosphatase 1B, PTP1B) activity in human Hela cells is carried out by including the following steps.

[0050] (1) The synthetic substrate sequence is: TPE-TPE-DADEpYL. The protected amino acids used and catalysts DIEA, HATU, DMAP, DCC and hexahydropyridine were purchased from Novabiochem, Acid Clear resin for synthesis was purchased from Peptide International, and DMF and DCM were purchased from Fisher. First, the resin is swelled with DMF for 10-60 minutes. At the same time, 100mg-1000mg of the first leucine was dissolved in anhydrous DCM, and then 10mg-250mg of DCC was added, and stirred in an ice bath for 1-30 minutes until a white precipitate formed. Evaporate the excess DCM with a rotary evaporator, dissolve it with 1-10mL DMF, and add it to the swollen resin to react for 10-60min...

Embodiment 3

[0054] First, synthesize TPE with the same method and process as in Example 1.

[0055] Secondly, in this embodiment, live cell imaging of protein phosphatase 2A (Protein phosphatase 2A, PP2A) activity in MCF-7 cells is performed by including the following steps.

[0056] (1) The synthetic substrate sequence is: TPE-RRREEEpTEEEAA. The protected amino acids used and catalysts DIEA, HATU, DMAP, DCC and hexahydropyridine were purchased from Novabiochem, Acid Clear resin for synthesis was purchased from Peptide International, and DMF and DCM were purchased from Fisher. First, the resin is swelled with DMF for 10-60 minutes. At the same time, 100mg-1000mg of the first alanine was dissolved in anhydrous DCM, and then 10mg-250mg of DCC was added, and stirred in an ice bath for 1-30 minutes until a white precipitate formed. Evaporate the excess DCM with a rotary evaporator, dissolve it with 1-10mL DMF, and add it to the swollen resin to react for 10-60min. Then wash the resin with ...

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Abstract

The invention discloses an enzyme-responsive self-aggregation luminous molecule and applications thereof in monitoring enzyme activity. The enzyme-responsive self-aggregation luminous molecule is formed by connecting a hydrolase substrate to an aggregation-induced luminous molecule, and can produce aggregation and induced luminescence after hydrolyzation under the action of the hydrolase. When the enzyme-responsive self-aggregation luminous molecule is added into living cells or extracellular reaction systems containing the corresponding hydrolase, the activity level of the hydrolase can be reflected through monitoring fluorescence signals by a fluorescence microscopic technology. The enzyme-responsive self-aggregation luminous molecule can be used for long-time space-time monitoring of the activity level of the hydrolase in cells.

Description

technical field [0001] The invention relates to the technical field of enzyme activity monitoring, in particular to an enzyme-responsive self-aggregation luminescent molecule and its application in monitoring enzyme activity. Background technique [0002] Any cell is composed of dynamic liposomes, proteins, nucleic acids, sugars and other macromolecules and small molecules with precisely adjustable structures and functions and ordered structures. Their complexity and variety are far beyond our current understanding. But as we all know, the state of cells directly reflects the state of organisms, so human beings must try various ways to know and understand the ubiquitous organisms, including ourselves. Understanding the interaction between molecules or molecular assemblies in cells is a way for us to recognize cells, which not only helps us understand life phenomena, but also helps us better diagnose and correct ourselves. With the rapid development of computer simulation t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
Inventor 梁兴杰张旭邹国漳赵元元
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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