β-lactamase substrates and methods of their use for the diagnosis of tuberculosis

Patent metadata

Field	Value
Publication number	US-9677112-B2
Application number	US-201314401775-A
Country	US
Kind code	B2
Filing date	May 15, 2013
Priority date	May 15, 2012
Publication date	Jun 13, 2017
Grant date	Jun 13, 2017

Abstract

β-Lactamase substrates and methods for using the substrates to detect β-lactamase diagnose tuberculosis.

First claim

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A compound having the formula:

or an ester or a salt thereof,
wherein
A is selected from the group consisting of:
(a) substituted and unsubstituted C6-C10 aryl; and
(b) substituted and unsubstituted C3-C7 heteroaryl;

R 1 is selected from the group consisting of:
(a) methoxy; and
(b) ethoxy;

R 2 is selected from the group consisting of hydrogen, C1-C3 alkyl, C1-C3 alkyl substituted with one or more halogens, and substituted piperazine;
X is O or S;
R a , R b , R c , and R d are independently selected from the group consisting of hydrogen, halogen, nitro, C1-C3 alkyl, and C1-C3 alkyl substituted with one or more halogens;
Z is a moiety that provides a fluorescent, luminescent, or colorimetric signal when released from the compound; and
n is 0 or 1.

2. The compound of claim 1 , wherein A is phenyl.

3. The compound of claim 1 , wherein R 1 is methoxy.

4. The compound of claim 1 , wherein X is O.

5. The compound of claim 1 , wherein n is 0.

6. The compound of claim 1 , wherein n is 1.

7. The compound of claim 1 , wherein Z is a fluorescent moiety.

8. The compound of claim 1 , wherein Z is a fluorescent phenolic dye moiety.

9. The compound of claim 1 , wherein Z is selected from the group consisting of a courmarin moiety, a xanthene moiety, a resorufin moiety, a cyanine moiety, a difluoroboradiazaindacene moiety, a bimane moiety, an acridine moiety, an isoindole moiety, a dansyl moiety, an aminophthalic hydrazide moiety, an aminophthalimide moiety, an aminonaphthalimide moiety, a quinine moiety, a dicyanovinyl moiety, a tricyanovinyl moiety, an indolaniline moiety, an indamine moiety, and derivatives thereof.

10. The compound of claim 1 , wherein Z is a xanthene moiety selected from the group consisting of a fluorescein moiety, a rhodol moiety, a rhodamine moiety, and derivatives thereof.

11. The compound of claim 1 , wherein Z is

12. The compound of claim 1 , wherein Z is

wherein R′ is hydrogen or aryl.

13. The compound of claim 12 , wherein R′ is phenyl or substituted phenyl.

14. The compound of claim 1 , wherein Z is

15. A compound of claim 1 having the formula:

or an ester or a salt thereof, wherein R 1 is methoxy.

16. A method for detecting beta-lactamase in a sample, comprising:
(a) contacting a sample with a compound of claim 1 ; and
(b) measuring an optical signal generated from contacting the sample with the compound.

17. A method for diagnosing tuberculosis, comprising:
(a) contacting a sample with a compound of claim 1 ; and
(b) measuring an optical signal generated from contacting the sample with the compound.

18. The method of claim 17 , wherein the sample is sputum, pleural fluid, spinal fluid, blood, urine, saliva, stool, tissue biopsies, tissue homogenates, directly in live animals or human patients, or a sample obtained by swabbing an area of interest on a subject.

19. The method of claim 17 , wherein the sample comprises a pathogenic bacterial species selected from Bacteroides, Clostridium, Streptococcus, Staphylococcus, Pseudomonas, Haemophilus, Legionella, Mycobacterium, Escherichia, Salmonella, Shigella , or Listeria.

20. The method of claim 17 , wherein measuring an optical signal comprises measuring fluorescence emission intensity.

21. The method of claim 17 , wherein measuring an optical signal comprises measuring absorbance intensity.

22. The method of claim 17 , wherein measuring an optical signal comprises measuring luminescence emission intensity.

Assignees

• Texas A & M Univ Sys
• Univ Leland Stanford Junior

Inventors

• Rao Jianghong
• Cirillo Jeffrey D
• Xie Hexin
• Sacchettini James C

Classifications

• C07D501/26

  Methylene radicals, substituted by oxygen atoms; Lactones thereof with the 2-carboxyl group · CPC title

• C12Q1/34 Primary

  involving hydrolase · CPC title

• C07D501/24 Primary

  with hydrocarbon radicals, substituted by hetero atoms or hetero rings, attached in position 3 · CPC title

• C07D501/30

  with the 7-amino-radical acylated by an araliphatic carboxylic acid · CPC title

• A61P31/06

  for tuberculosis · CPC title