6,7-trans cephalosporin-based probes for detecting bacteria expressing a metallo-beta-lactamase
View Patent ↗The present invention discloses a probe useful for the selective detection of metallo-beta-lactamases, in particular carbapenemases, thereby distinguishing those species of bacteria that are carbapenem-resistant from bacterial strains that are sensitive. Cephalospori based probes that have the 6,7 R,R configuration are susceptible to cleavage by beta-lactamases but cannot distinguish between cleavage by metallo-beta-lactamases and other beta-lactamases. By modifying a side group of the cephalosporin, selectivity can be introduced allowing the probes to distinguish between various types of metallo-beta-lactamases.
1. A probe selectively cleavable by a metallo-β-lactamase, wherein said probe consists of an isolated cephalosporin enantiomer having a 6,7-trans configuration and a detectable label attached thereto, wherein the probe has the formula I or formula II:
wherein:
R 1 is selected from the group consisting of: methyl, ethyl, isopropyl, tert-butyl, benzyl-, and tosyl, and wherein the detectable label is (2-oxo-2H-chromen-7-yl) (III) or (4S)-2-(6-oxo-1,3-benzothiazol-2-yl)-4,5-dihydrothiazole-4-carboxylic acid (IV) having the structures:
2. The composition of claim 1 , wherein the probe is selected from the group consisting of:
3. A method of selectively detecting a metallo-β-lactamase activity, said method comprising contacting a sample suspected of having a metallo-β-lactamase activity with a probe selectively cleavable by a metallo-β-lactamase, wherein said probe consists of an isolated cephalosporin enantiomer having a 6,7-trans configuration and a detectable label attached thereto, wherein the probe has the formula I or formula II:
wherein:
R 1 is selected from the group consisting of: methyl, ethyl, isopropyl, tert-butyl, benzyl-, and tosyl, and wherein the detectable label is (2-oxo-2H-chromen-7-yl) (III) or (4S)-2-(6-oxo-1,3-benzothiazol-2-yl)-4,5-dihydrothiazole-4-carboxylic acid (IV) having the structures:
allowing an effective period for a metallo-β-lactamase activity to cleave the cephalosporin moiety, thereby releasing the detectable label from the cephalosporin moiety, whereby a detectable signal is generated; and detecting the signal, wherein a detectable signal indicates that the sample contains a metallo-β-lactamase activity or a source of a metallo-β-lactamase activity.
4. The method of claim 3 , wherein the probe is selected from the group consisting of:
5. The method of claim 3 , wherein the sample is a bacterial culture.
6. The method of claim 3 , wherein the sample has been isolated from an animal or human subject.
7. The method of claim 5 , wherein the sample is a bacterial culture generated from a sample from a human or animal subject.