Environmental sensitive fluorogenic compounds and their application for singlet oxygen and protein detection

• US 8,173,698 B2
• Filed: 02/29/2008
• Issued: 05/08/2012
• Est. Priority Date: 11/19/2003
• Status: Expired due to Fees

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Johnson, Iain, Liu, Jixiang, Patch, Brian, Leung, Wai-Yee
• Primary Examiner(s): AULAKH, CHARANJIT
• Application Number:
  US12/040,850
• Publication Number:
  US 20080160627A1
• Time in Patent Office: 1,530 Days
• Field of Search: 514/454, 514/452, 549/391, 549/358, 436/800
• US Class Current: 514/454
• CPC Class Codes: C09B 1/00 Dyes with anthracene nucleus not condensed with any other ring
• CPC Class Codes: C09B 11/08 Phthaleins; Phenolphthaleins; Fluorescein
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C09B 19/00 Oxazine dyes
• CPC Class Codes: C09B 57/02 Coumarine dyes
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10T 436/20 Oxygen containing

Abstract

The present invention provides fluorogenic probes and corresponding fluorescent compounds, methods of using the probes, compounds and kits that include the probes.

Claims

1. A fluorogenic compound, where the compound is:

2. A method for the determination of the presence or absence of singlet oxygen in a sample, wherein the method comprises:

a) contacting the sample with a fluorogenic compound according to claim 1 to prepare a labeled sample;

b) incubating the labeled sample for a sufficient amount of time to prepare an incubated sample, wherein the singlet oxygen reacts with the fluorogenic compound to produce a fluorescent product;

c) illuminating the incubated sample with an appropriate wavelength to prepare an illuminated sample; and

d) observing the illuminated sample whereby the presence or absence of the singlet oxygen in the sample is determined.

3. The method according to claim 2, wherein the singlet oxygen is a product formed by a photosensitizer and ground state oxygen in the presence of visible light, hypochlorites and hydrogen peroxide, benzyl cyanide and hydrogen peroxide, oxalyl chloride and hydrogen peroxide, sodium molybdate and hydrogen peroxide, ozone and phosphines, ozone and phosphites, electrodeless discharge of ground state oxygen, hydrogen peroxide in an aqueous solution with Fe (II), (III), or Ce (IV) ions, or pyrolysis of aromatic endoperoxides.

4. The method according to claim 2, wherein the singlet oxygen is a product formed by the reaction of a photosensitizer and ground state oxygen in the presence of visible light.

5. The method according to claim 3, wherein the photosensitizer is an aromatic carbonyl compound, condensed aromatic compound, acridine dye, coumarin dye, crystal violet, fluorene derivative, naphthalocyanine, porphyrin derivative, chlorin, phthalocyanine, thiazine dye, thioketone, xanthene dye, platinum (II) terpyridyl acetylide complex, or combinations thereof.

6. The method according to claim 3, wherein the photosensitizer is an acetonapthone, acetophenone, benzophenone, acridine orange, eosin, fluorene, fluorenone, fluorescein, methylene blue, naphthalocyanine, phthalocyanine, rose bengal, 5,10,15,20-tetrakis(m-hydroxyphenyl)chlorin, thiocoumarin, toluidine blue, methylene blue, zinc tetraphenylporphyrin tetrasulfonate, or combinations thereof.

7. The method according to claim 3, wherein the photosensitizer is methylene blue.

8. The method according to claim 2, wherein the singlet oxygen is a product formed by the reaction of sodium molybdate and hydrogen peroxide.

9. A kit for the determination of singlet oxygen in a sample, wherein the kit comprises:

a) a reaction buffer; and

,

b) a fluorogenic compound according to claim 1.

10. The kit according to claim 9, further comprising instructions for use of the kit.

11. The kit according to claim 9, further comprising a singlet oxygen generator.

12. The kit according to claim 9, further comprising methylene blue, sodium molybdate or hydrogen peroxide.

13. A composition comprising:

a) a sample; and

b) a fluorogenic compound according to claim 1.

14. The composition according to claim 13, further comprising a singlet oxygen generator in an amount sufficient to produce singlet oxygen.

15. The composition according to claim 13, further comprising methylene blue, sodium molybdate or hydrogen peroxide.