Fluorescent probe, singlet oxygen detection agent, and singlet oxygen detection method

Patent metadata

Field	Value
Publication number	US-10564164-B2
Application number	US-201515320162-A
Country	US
Kind code	B2
Filing date	Jun 17, 2015
Priority date	Jun 17, 2014
Publication date	Feb 18, 2020
Grant date	Feb 18, 2020

Abstract

An object of the present invention is to provide a fluorescent probe that easily penetrates a cell and is capable of selectively and efficiently detecting singlet oxygen generated in the cell, a singlet oxygen detection agent containing the fluorescent probe, and a singlet oxygen detection method using the fluorescent probe. The fluorescent probe for detecting singlet oxygen comprises a compound having a silicon-rhodamine skeleton and a skeleton that increases the fluorescence of the fluorescent probe after trapping singlet oxygen to be greater than the fluorescence before trapping singlet oxygen, or a salt, hydrate, or solvate thereof.

First claim

We claim:

1. A fluorescent probe for detecting singlet oxygen, comprising a compound represented by Formula (1):

wherein R 1 to R 4 are the same or different, and each represents hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, or substituted or unsubstituted aryl; R 5 and R 6 are the same or different, and each represents hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, or substituted or unsubstituted aryl; R 7 and R 8 are the same or different, and each represents hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted alkoxy, or acid addition salt; R 9 and R 10 are the same or different, and each represents hydroxy, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, or acid addition salt; n is an integer of 0 to 3; and m is an integer of 0 to 4.

2. The fluorescent probe according to claim 1 , wherein the compound is represented by Formula (1A):

wherein R 1 to R 10 , n, and m are as defined in claim 1 .

3. The fluorescent probe according to claim 1
wherein each of R 1 to R 8 is a methyl, and n and m are both 0.

4. A singlet oxygen detection agent, comprising the fluorescent probe according to claim 1 .

5. The singlet oxygen detection agent according to claim 4 , wherein the singlet oxygen detection agent contains 10 to 500 nmol/L of the fluorescent probe.

6. A method for detecting singlet oxygen generated in a cell, comprising the steps of:
(1) culturing a cell under a condition in which singlet oxygen is generated;
(2) preparing and culturing a mixed culture solution containing the culture solution obtained in step (1) and the fluorescent probe of claim 1 , wherein the concentration of the fluorescent probe is 10 to 500 nmol/L; and
(3) irradiating the mixed culture solution obtained in step (2) with light.

7. The method for detecting singlet oxygen according to claim 6 , wherein the condition in which singlet oxygen is generated in step (1) is a condition having a photosensitizes.

8. The method for detecting singlet oxygen according to claim 7 , wherein the photosensitizer is protoporphyrin IX.

9. The method for detecting singlet oxygen according to claim 6 , wherein the cell is a cancer cell.

10. The method for detecting singlet oxygen according to claim 6 , wherein the method site-selectively detects singlet oxygen generated near a mitochondrial inner membrane in the cell.

11. A compound represented by Formula (1):

wherein R 1 to R 4 are the same or different, and each represents hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, or substituted or unsubstituted aryl; R 5 and R 6 are the same or different, and each represents hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, or substituted or unsubstituted aryl; R 7 and R 8 are the same or different, and each represents hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted alkoxy, or acid addition salt; R 9 and R 10 are the same or different, and each represents, hydroxy, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aryl, or acid addition salt; n is an integer of 0 to 3; and m is an integer of 0 to 4.

12. The compound according to claim 11 , wherein the compound is represented by Formula (1A):

wherein R 1 to R 10 , n, and m are as defined in claim 11 .

13. A cell assay reagent, comprising the fluorescent probe of claim 1 .

14. The singlet oxygen detection agent according to claim 4 , which is a singlet oxygen detection agent generated near a mitochondrial inner membrane.

15. The singlet oxygen detection agent according to claim 4 , which is color rendering agent of mitochondria.

16. The compound according to claim 11 , wherein each of R 1 to R 8 is a methyl, and n and m are both 0.

17. A cell assay reagent, comprising the singlet oxygen detection agent of claim 4 .

18. A cell assay reagent, comprising the compound of claim 11 .

Assignees

• Univ Osaka

Inventors

• Majima Tetsuro
• Kim Sooyeon
• Tachikawa Takashi
• Fujitsuka Mamoru

Classifications

• C07F7/10

  containing nitrogen {having a Si-N linkage} · CPC title

• C07F7/0816

  said ring comprising Si as a ring atom · CPC title

• G01N33/582 Primary

  with fluorescent label · CPC title