Luminescent probe and its preparation method and application

Patent metadata

Field	Value
Publication number	US-12305098-B2
Application number	US-202418768028-A
Country	US
Kind code	B2
Filing date	Jul 10, 2024
Priority date	Aug 24, 2023
Publication date	May 20, 2025
Grant date	May 20, 2025

Abstract

A luminescent probe and its preparation method and application are provided. The luminescent probe has a steric hindrance group R1 of aliphatic hydrocarbon structure such as adamantane or norborneol, a detection group R2 of nitrobenzyl and its derivative structure, an electron-withdrawing group R3 containing cyano group and an electron-donating group methoxy group. In the presence of HSA or BSA, the detection group is cut off to form a parent structure that exposes atomic oxygen anions and is activated under external light irradiation, the luminescent probe can be used in solution or cells, when detecting HSA or BSA, the luminescent probe has obvious chemiluminescence characteristics, which can sensitively distinguish HSA and BSA, quantitatively analyze HSA and BSA, and determine the mixing ratio of HSA and BSA at the same time, and the luminescent probe has been successfully used for cell fluorescence imaging.

First claim

What is claimed is:

1. A luminescent probe, wherein the luminescent probe has a structural formula shown in formula (I)

wherein R 1 is selected from one of

R 2 is selected from one of

and R 3 is selected from one of

2. The luminescent probe according to claim 1 , wherein the structural formula of the luminescent probe is

3. A preparation method for a luminescent probe, wherein the luminescent probe has a structural formula shown in formula (I)

wherein R 1 is selected from one of

R 2 is selected from one of

and R 3 is selected from one of

the preparation method comprising the following steps:
1) mixing 4-(dicyanomethylene)-2, 6-dimethyl-4H-pyran, a first compound, acetonitrile, and piperidine, then performing a first reaction, to obtain a phenolic hydroxyl precursor;
2) mixing the phenolic hydroxyl precursor, a second compound, and a solvent, then cooling to 0° C., adding nitro compounds, and performing a second reaction, to obtain the luminescent probe with the structural formula (II), (III) or (IV);
in step 1), the first compound is

and
in step 2), the second compound is triethylamine or cesium carbonate; the nitro compounds are benzyl 4-nitrochloroformate or 4-nitrobenzyl bromide.

4. The preparation method according to claim 3 , wherein in step 1), a molar volume ratio of the 4-(dicyanomethylene)-2, the 6-dimethyl-4H-pyran, the first compound, the acetonitrile, and the piperidine is 0.6-2.5 mmol:0.6-2.5 mmol:20-30 mL:0.1-1 mL.

5. The preparation method according to claim 3 , wherein in step 1), a reaction temperature is 20-30° C., and a reaction time is 5-7 h.

6. The preparation method according to claim 5 , wherein in step 2), a molar volume ratio of the phenolic hydroxyl precursor, the second compound, the solvent, and the nitro compounds is 0.06-0.12 mol:0.1-0.2 mmol:15-25 mL:0.1-0.2 mmol.

7. The preparation method according to claim 4 , wherein in step 2), a reaction temperature is 0-30° C., and a reaction time is 20-26 h.

8. An application of a luminescent probe in a detection of human serum albumin and bovine serum albumin, wherein the luminescent probe has a structural formula shown in formula (I)

wherein R 1 is selected from one of

R 2 is selected from one of

and R 3 is selected from one of

wherein the application comprising the following steps: mixing a luminescent probe stock solution with a mixed solvent and a serum albumin stock solution in turn, afterwards, performing an incubation and detection in turn.

9. The application according to claim 8 , wherein a concentration of the luminescent probe stock solution is 0.8×10 31 3 −1.5×10 −3 M; the mixed solvent is prepared from dimethyl sulfoxide and a phosphate buffered saline (PBS) buffer solution, wherein a volume ratio of the dimethyl sulfoxide and the PBS buffer solution is 1:2-3, a pH value of the PBS buffer solution is 7.0-7.5; and a concentration of the serum albumin stock solution is 2.0×10 −4 −1.0×10 −2 M.

10. The application according to claim 8 , wherein a volume ratio of the luminescent probe stock solution, the mixed solvent, and the serum albumin stock solution is 20 μL:1.84-2 mL:0-160 μL; and an incubation temperature is 36.5-37.5° C., an incubation time is 42-46 min.

11. The preparation method according to claim 4 , wherein in step 1), a reaction temperature is 20-30° C., and a reaction time is 5-7 h.

12. The preparation method according to claim 6 , wherein in step 2), a reaction temperature is 0-30° C., and a reaction time is 20-26 h.

13. The application according to claim 8 , wherein in the luminescent probe, the structural formula of the luminescent probe is

14. The application according to claim 9 , wherein a volume ratio of the luminescent probe stock solution, the mixed solvent, and the serum albumin stock solution is 20 μL:1.84-2 mL:0-160 μL; and an incubation temperature is 36.5-37.5° C., an incubation time is 42-46 min.

Assignees

• Univ East China Science & Tech

Inventors

• Guo Zhiqian
• Li Juan
• Zhang Yutao
• Lu Yao
• Yan Chenxu
• Zhao Meiling
• Zhao xiuyan
• Zhu Weihong

Classifications

• G01N2333/765

  Serum albumin, e.g. HSA · CPC title

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• C09K2211/1088

  containing oxygen as the only heteroatom · CPC title

• C07D407/10

  linked by a carbon chain containing aromatic rings · CPC title

• C07D309/34 Primary

  having three or more double bonds between ring members or between ring members and non-ring members · CPC title