HYDROGEN PEROXIDE SENSORS BASED UPON PHOTO-INDUCED ELECTRON TRANSFER

• US 20120183984A1
• Filed: 07/22/2010
• Published: 07/19/2012
• Est. Priority Date: 07/22/2009
• Status: Active Grant

• Current Assignee: OPTI Medical Systems, Inc. (IDEXX Laboratories Incorporated)
• Original Assignee: OPTI Medical Systems, Inc. (IDEXX Laboratories Incorporated)
• Inventors: He, Huarui, Lin, Chao
• Granted Patent: US 9,018,377 B2
• Time in Patent Office: Days
• US Class Current: 435/28
• CPC Class Codes: C07D 221/14 Aza-phenalenes, e.g. 1,8-naphthalimide
• CPC Class Codes: C07D 401/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: Y10T 436/206664 Ozone or peroxide

Abstract

The invention provides compounds of formula I F-L-Q (I) where F comprises a fluorophore capable of absorbing energy at an excitation wavelength and, in the absence of a quencher, emitting energy at an emission wavelength, which is different than the excitation wavelength; Q comprises a quencher; L comprises a linker moiety having two ends, one end being covalently bound to F and the other end being covalently bound to Q. The compounds are capable of undergoing a reversible reaction (1), provided below: (1) where Q⁺ is an oxidized form of Q representing the absence of a quencher, Ox comprises an oxidizing agent, which is capable of oxidizing Q to its oxidized form Q, and Red comprises a reducing agent, which is capable of converting Q back to its reduced form Q. The compounds can undergo photo-induced electron transfer when irradiated with energy and when Q exists in its oxidized form, Q⁺. The invention also provides methods of detecting and determining the presence of analytes and/or hydrogen peroxide in a sample, as well as a substrate that comprises the compound of formula I.

Claims

1. A compound having a chemical structure conforming to formula (I) below, or a salt thereof:

F-L-Q 

 

(I)

in which

F comprises a fluorophore capable of absorbing energy at an excitation wavelength and, in the absence of a quencher, emitting energy at an emission wavelength, which is different than the excitation wavelength;

Q comprises a quencher;

L comprises a linker moiety having two ends, one end being covalently bound to F and the other end being covalently bound to Q;

said compound capable of undergoing a reversible reaction (1), provided below;

2. The compound of claim 1 in which Q is capable of absorbing energy from F via a photo-induced electron transfer (PET) quenching mechanism in the absence of Ox, and in which Q⁺ is incapable of absorbing energy from F via a PET quenching mechanism.

3. The compound of claim 1 in which Q comprises a five- or six-membered aryl or heteroaryl group that is substituted by at least one substituent selected from the group consisting of OR¹, SR¹, and N(R¹)₂, wherein each instance of R¹ is independently selected from the group consisting of hydrogen and C_1-6-alkyl.

4. The compound of claim 1 in which L comprises a saturated or unsaturated, linear or branched aliphatic chain including 1-6 carbon atoms, wherein 1-3 carbon atoms are optionally and independently replaced by —

NR¹—

, —

O—

, or —

S—

, wherein R¹ is hydrogen or C_1-6-alkyl.

5. The compound of claim 1, or a salt thereof, in which the compound has a chemical structure that conforms to formula IA, below:

6. The compound of claim 5 in which:

R² is selected from C_1-8-alkyl, C_1-8-alkoxy, C_1-8-alkyl-aryl, C_1-8-alkyl-heteroaryl, C_1-8-alkyl-C_1-8-alkoxy;

X and Y are independently selected from the group consisting of —

NR¹— and

—

O—

;

m is 1 or 2; and

n is 2 or 3.

7. The compound of claim 5 which is selected from the group consisting of:

8. The compound of claim 7 which is selected from the group consisting of:

9. The compound of claim 8 which is:

10. The compound of claim 8 which is:

11. A method of determining a concentration of an analyte in a sample comprising:

(a) contacting a sample suspected of containing an analyte with

(i) a sufficient amount of a compound having a chemical structure conforming to formula (I) below, or a salt thereof;

F-L-Q 

 

(I)

in which

F comprises a fluorophore capable of absorbing energy at an excitation wavelength and, in the absence of a quencher, emitting energy at an emission wavelength, which is different than the excitation wavelength;

Q comprises a quencher;

L comprises a linker moiety having two ends, one end being covalently bound to F and the other end being covalently bound to Q;

said compound capable of undergoing a reversible reaction (1), provided below;

12. The method of claim 11 in which Q is capable of absorbing energy from F via a photo-induced electron transfer (PET) quenching mechanism in the absence of Ox, and in which Q⁺ is incapable of absorbing energy from F via a PET quenching mechanism.

13. A substrate useful in the detection of an analyte in a sample comprising a surface onto which is affixed

(i) an analytically effective amount of a compound having a chemical structure conforming to formula (I) below, or a salt thereof:

F-L-Q 

 

(I)

in which

F comprises a fluorophore capable of absorbing energy at an excitation wavelength and, in the absence of a quencher, emitting energy at an emission wavelength, which is different than the excitation wavelength;

Q comprises a quencher;

L comprises a linker moiety having two ends, one end being covalently bound to F and the other end being covalently bound to Q;

said compound capable of undergoing a reversible reaction (1), provided below;

14. The substrate of claim 13 in which Q is capable of absorbing energy from F via a photo-induced electron transfer (PET) quenching mechanism in the absence of Ox, and in which Q⁺ is incapable of absorbing energy from F via a PET quenching mechanism.

15. The substrate of claim 13 for determining a concentration of the analyte in the sample.

16. A method of determining a concentration of hydrogen peroxide in a sample comprising:

(a) contacting a sample suspected of containing hydrogen peroxide (H₂O₂) with a sufficient amount of a compound having a chemical structure conforming to formula (I) below, or a salt thereof;

F-L-Q 

 

(I)

in which

F comprises a fluorophore capable of absorbing energy at an excitation wavelength and, in the absence of a quencher, emitting energy at an emission wavelength, which is different than the excitation wavelength;

Q comprises a quencher;

L comprises a linker moiety having two ends, one end being covalently bound to F and the other end being covalently bound to Q;

said compound capable of undergoing a reversible reaction (1), provided below;

17. The method of claim 16 in which Q is capable of absorbing energy from F via a photo-induced electron transfer (PET) quenching mechanism in the absence of Ox, and in which Q⁺ is incapable of absorbing energy from F via a PET quenching mechanism

18. A method of detecting the presence of hydrogen peroxide in a sample comprising:

(a) contacting a sample suspected of containing hydrogen peroxide with a sufficient amount of a compound having a chemical structure conforming to formula (I) below, or a salt thereof;

F-L-Q 

 

(I)

in which

F comprises a fluorophore capable of absorbing energy at an excitation wavelength and, in the absence of a quencher, emitting energy at an emission wavelength, which is different than the excitation wavelength;

Q comprises a quencher;

L comprises a linker moiety having two ends, one end being covalently bound to F and the other end being covalently bound to Q;

said compound capable of undergoing a reversible reaction (1), provided below;

19. The method of claim 18 in which Q is capable of absorbing energy from F via a photo-induced electron transfer (PET) quenching mechanism in the absence of Ox, and in which Q⁺ is incapable of absorbing energy from F via a PET quenching mechanism.

20. The method of claim 18 which further comprises measuring an intensity of the emitted energy at the emission wavelength.