Ratiometric fluorescent probe for detecting aminopeptidase N, and preparation method and use thereof

Patent metadata

Field	Value
Publication number	US-12480948-B2
Application number	US-202217856052-A
Country	US
Kind code	B2
Filing date	Jul 1, 2022
Priority date	Apr 25, 2022
Publication date	Nov 25, 2025
Grant date	Nov 25, 2025

Abstract

Provided is a ratiometric fluorescent probe for detecting aminopeptidase N, and a preparation method and use thereof. In the present disclosure, a Nile blue derivative is adopted as a fluorophore and alanyl is adopted as an identification unit to design and synthesize the ratiometric fluorescent probe NB-APN for detecting APN. After the probe reacts with APN, the NB blocked by alanyl is released, resulting in an increase of a fluorescence peak at 675 nm and a decrease of a fluorescence peak at 610 nm. A ratio signal of the probe exhibits a sensitive response to APN, with a detection limit as low as 15 pg/mL. The probe can be used for quantitative detection of APN in a diluted urine sample, and can also be used for ratiometric fluorescent imaging of APN in a cell model and in vivo fluorescent imaging of APN in a nude mouse tumor model.

First claim

What is claimed is:

1 . A ratiometric fluorescent probe for detecting aminopeptidase N, wherein a structure of the ratiometric fluorescent probe comprises an alanyl group and a Nile blue derivative;
wherein the Nile blue derivative has a structural formula as follows:

wherein the ratiometric fluorescent probe has a structural formula as follows:

2 . The ratiometric fluorescent probe according to claim 1 , wherein in the structure of the ratiometric fluorescent probe, a quantity ratio of the alanyl group to the Nile blue derivative is 1:1.

3 . A kit for fluorometric analysis of aminopeptidase N in a body fluid or fluorescent imaging of aminopeptidase N in a cell comprising the ratiometric fluorescent probe according to claim 1 .

4 . The kit for fluorescent imaging of aminopeptidase N in a cell according to claim 3 , wherein the cell comprises a human normal cell and a human tumor cell.

5 . The kit for fluorometric analysis of aminopeptidase N in a body fluid according to claim 3 , wherein in the structure of the ratiometric fluorescent probe, a quantity ratio of the alanyl group to the Nile blue derivative is 1:1.

Assignees

• Univ Jilin

Inventors

• Song Daqian
• Ma Pinyi
• Gao Dejiang

Classifications

• G01N2333/948

  acting on peptide bonds (3.4) · CPC title

• G01N33/582

  with fluorescent label · CPC title

• G01N21/6456

  Spatial resolved fluorescence measurements; Imaging · CPC title

• G01N33/573 Primary

  for enzymes or isoenzymes · CPC title

• C09K2211/1033

  with oxygen · CPC title