Self-immolative probes for enzyme activity detection

Patent metadata

Field	Value
Publication number	US-12497645-B2
Application number	US-202318161610-A
Country	US
Kind code	B2
Filing date	Jan 30, 2023
Priority date	Nov 16, 2010
Publication date	Dec 16, 2025
Grant date	Dec 16, 2025

Abstract

Provided is a compound comprising the structure:

(SIG)-(SI-MOD) m .

In this compound, SIG is a signaling molecule, SI is a self-immolative structure bound to SIG such that SIG has a reduced signal relative to the signal of SIG without SI, MOD is a moiety bound to SI that is subject to modification by an activator, and m is an integer from 1 to about 10. With this compound, when MOD is modified by an activator, SI is destabilized and self-cleaved from SIG such that SIG generates an increased signal. Also provided is a method of determining whether a sample comprises an activator, using the above-described compound. Additionally provided is a method of determining whether a cell comprises a nitroreductase using the above-described compound where nitroreductase is the activator. Further provided is a method of determining whether a mammalian cell is hypoxic using the above-described compound where nitroreductase is the activator. A method of detecting a microorganism that comprises a nitroreductase, using the above-described compound where nitroreductase is the activator, is also provided. Also provided is a method of identifying nitroreductase in a sample, using the above-described compound where nitroreductase is the activator.

First claim

What is claimed is:

1 . A compound comprising the structure:
(SIG)-(SI-MOD) m wherein SIG is a signaling molecule, SI is a self-immolative structure bound to SIG such that SIG has a reduced signal relative to the signal of SIG without SI, MOD is a moiety bound to SI that is subject to modification by an activator, and m is an integer from 1 to about 10, wherein, when MOD is modified by an activator, SI is destabilized and self-cleaved from SIG such that SIG generates an increased signal; wherein the compound further comprises the structure of

wherein
S—S is MOD;
each R 2 is independently a hydrogen, an unsubstituted straight-chain, branched or cyclic alkyl, alkenyl or alkynyl group, a substituted straight-chain, branched or cyclic alkyl, alkenyl or alkynyl group wherein one or more C, CH or CH 2 groups are substituted with an O atom, N atom, S atom, NH group, CO group, OCO group or CONR 3 group, or an unsubstituted or substituted aromatic group; and
q is an integer from 1 to 4.

2 . The compound of claim 1 , wherein the activator is a disulfide reducing agent.

3 . The compound of claim 2 , wherein the disulfide reducing agent is glutathione, cysteine, or homocysteine.

4 . The compound of claim 2 , wherein the disulfide reducing agent is glutathione.

5 . The compound of claim 1 , wherein m>1.

6 . The compound of claim 1 , wherein m=1.

Assignees

• Enzo Life Sciences Inc

Inventors

• Lebedeva Irina
• Patton Wayne Forrest
• Szczepanik Maciej
• Xiang Yuejun
• Pande Praveen

Classifications

• C09K2211/1088

  containing oxygen as the only heteroatom · CPC title

• C09K2211/1022

  bridged by heteroatoms, e.g. N, P, Si or B · CPC title

• C07D493/10

  Spiro-condensed systems · CPC title

• C07D491/20

  Spiro-condensed systems · CPC title

• C07D491/052

  the oxygen-containing ring being six-membered · CPC title