Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Aggregation-induced emission fluorescent probe for detecting aminopeptidase N and preparation of aggregation-induced emission fluorescent probe

A technology of aggregation-induced emission and fluorescent probes, which is applied in the field of fluorescent probes to achieve the effects of fast response, low interference, good sensitivity and selectivity

Active Publication Date: 2021-08-10
UNIV OF JINAN
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to the best of our knowledge, an activatable aggregation-induced emission-based fluorescent probe for the detection of APN with long emission wavelengths has hardly been reported.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Aggregation-induced emission fluorescent probe for detecting aminopeptidase N and preparation of aggregation-induced emission fluorescent probe
  • Aggregation-induced emission fluorescent probe for detecting aminopeptidase N and preparation of aggregation-induced emission fluorescent probe
  • Aggregation-induced emission fluorescent probe for detecting aminopeptidase N and preparation of aggregation-induced emission fluorescent probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A method for preparing an aggregation-induced emission fluorescent probe for detecting aminopeptidase N, the steps comprising:

[0025] 1) Synthesis of Compound 1:

[0026] in N 2 Under atmosphere, 2-methylquinoline (1.79 g, 12.5 mmol) and iodomethane (5.50 g, 35 mmol) were dissolved in acetonitrile (25 mL), and the reaction solution was refluxed for 10 hours, and the solvent was evaporated under reduced pressure to obtain Compound 1, yield 75%.

[0027] Synthesis of compound 2:

[0028] Compound 1 (1.74 g, 6 mmol) and malononitrile (1.20 g, 18 mmol) were dissolved in absolute ethanol (15 mL), stirred continuously at 0°C, and reacted for 6 hours after adding sodium ethoxide, and the mixture was Pour into ice water, adjust the pH to 8, filter the resulting precipitate, wash with water and vacuum dry to obtain compound 2 with a yield of 85%.

[0029] Synthesis of compound 3:

[0030] Compound 2 (1.40 g, 7.5 mmol) and p-hydroxybenzaldehyde (1.65 g, 13.5 mmol) were dis...

Embodiment 2

[0038] Measurement of Absorption Spectrum and Fluorescence Spectrum of Probe MQ-N

[0039] Take the MQ-N synthesized in Example 1 in a test tube to prepare a stock solution (1.0 mM) of the probe MQ-N in DMSO. Preparation of NaCl, KCl, CaCl in distilled water 2 , ZnCl 2 , MgCl 2 , CuCl 2 , H 2 o 2, NaClO, glucose, GSH, Cys, Hcy, alkaline phosphatase, γ-glutamyl transpeptidase, nitroreductase, carboxylesterase and other stock solutions. All the above stock solutions were adjusted to a final volume of 5 mL with phosphate buffered saline. For spectroscopic experiments, after incubating the MQ-N stock solution with a certain amount of analyte in a quartz cuvette at pH = 7.4 for 30 min, transfer 3 mL of the reaction solution to a 1 cm quartz cuvette, and set the excitation to 480 nm, the emission wavelength is 500-700 nm. Meanwhile, a control group without aminopeptidase N was prepared and compared under the same conditions. MQ-N showed double absorption peaks around 350 nm...

Embodiment 3

[0041] Evaluate the effect of different pH and temperature on the fluorescence of probe MQ-N

[0042] Taking the MQ-N stock solution (10 μM) in Example 2, the probe is almost non-fluorescent when the pH range is 4.0-8.0 and the temperature range is 23-40°C, which shows that the probe has high stability . After reaction with aminopeptidase N, the probe produced a significant fluorescent response throughout the pH range tested, although the maximum fluorescence intensity occurred in the pH range of 6.0–7.4. On the other hand, the strongest fluorescence of the reaction solution occurs at about 37°C, which is consistent with the fact that enzymes generally have maximum activity at 37°C. The results indicated that the probe MQ-N has good function under normal physiological conditions.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an aggregation-induced emission fluorescent probe for detecting aminopeptidase N and a preparation method of the aggregation-induced emission fluorescent probe. The structure of a probe compound is shown as a formula I in the specification. The probe molecule is composed of three main parts: a quinoline-malononitrile-based aggregation-induced emission fluorophore, a self-cleavable linking group and an aminopeptidase N (APN) specific recognition group. When the N-terminal alanyl site of the probe compound is accurately hydrolyzed into amino by APN, the exposed amino is used as an electron donating group, and the electron push-pull effect of a conjugated system is promoted, so that the fluorescence is enhanced. The probe has the advantages that the response speed is high, the sensitivity is high, the Stokes shift is large, the emission wavelength is long, when the concentration of a solution is higher, the fluorescence emission is stronger, and the problem of aggregation-induced fluorescence quenching is effectively avoided.

Description

technical field [0001] The present invention relates to the in situ detection of the location and expression level of active enzymes in cells by small molecule fluorescent probes, more specifically the accurate detection of the content of endogenous aminopeptidase N by a fluorescent probe based on aggregation-induced emission, It belongs to the technical field of fluorescent probes. Background technique [0002] Aminopeptidase N (APN / CD13), known as membrane-bound zinc ion-dependent type II metalloprotease, can degrade neutral or basic amino acids at the N-terminal of protein polypeptide chains. APN accordingly participates in many important physiological and pathological processes in the body, such as signal transduction, immune regulation, hydrolysis of bioactive peptides, and the invasion process of coronaviruses. Many studies have shown that APN is highly expressed on the surface of most tumor cells compared with normal cells, and APN plays a crucial role in the growth,...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07D215/14C09K11/06A61K49/00
CPCC07D215/14C09K11/06A61K49/0032C09K2211/1007C09K2211/1014C09K2211/1029
Inventor 颜梅卫先哲张晶杨小凤朱彤李成芳于京华
Owner UNIV OF JINAN
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com