Activity-based probe compounds, compositions, and methods of use
View Patent ↗Activity-based probe compounds for use in labeling a cysteine protease are provided. The compounds are targeted to the protease through a specific targeting element. The compounds additionally include a detectable element, such as a fluorescent label, a radiolabel, or a chelator. In some cases, the compounds additionally include a quenching element that is released upon reaction with the protease. Also provided are compositions comprising the compounds and methods for using the compounds, for example in labeling a protease in an animal and in visualizing a tumor in an animal.
1. A compound for use in labeling a protease having the formula (II):
L 1 is a linker;
AA 1 is an amino acid side chain;
U is O, NH, or S;
R 1 is alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, or a protecting group, and is optionally substituted with 1 to 3 A groups;
each A is independently alkyl, alkenyl, alkynyl, alkoxy, alkanoyl, alkylamino, aryl, aryloxy, arylamino, aralkyl, aralkoxy, aralkanoyl, aralkamino, heteroaryl, heteroaryloxy, heteroarylamina, heteroaralkyl, heteroaralkoxy, heteroaralkanoyl, heteroaralkamino, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkoxy, cycloalkanoyl, cycloalkamino, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylalkyl, heterocyclylalkoxy, heterocyclylalkanoyl, heterocyclylalkamino, hydroxyl, thio, amino, alkanoylamino, aroylamino, aralkanoylamino, alkylcarboxy, carbonate, carbamate, guanidinyl, urea, halo, trihalomethyl, cyano, nitro, phosphoryl, sulfonyl, sulfonamido, or azido;
L 3 is a linker; and
Q comprises a quencher,
wherein D comprises a benzoindole dye having the structure:
wherein o is an integer from 1 to 4;
R 2 is a C 2 -C 8 alkyl group, substituted with a sulfonate or carbonate;
each R 3 is independently a C 1 -C 6 alkyl group; and
L 4 is an optionally substituted alkyl linker, wherein each carbon atom is optionally replaced with a heteroatom.
2. The compound of claim 1 , wherein the benzoindole dye has the structure:
3. The compound of claim 1 , wherein the benzoindole dye has the structure:
4. The compound of claim 1 , wherein L 1 is an optionally substituted alkyl linker, wherein each carbon atom is optionally replaced with a heteroatom.
5. The compound of claim 1 , wherein AA 1 is an aralkyl amino acid side chain, optionally substituted with 1 to 3 A groups.
6. The compound of claim 1 , wherein the U is O.
7. The compound of claim 1 , wherein L 3 is an optionally substituted alkyl linker, wherein each carbon atom is optionally replaced with a heteroatom.
8. The compound of claim 1 , wherein L 3 -Q is
wherein R comprises a QSY quencher or a QC-1 quencher; and
n is an integer from 1-8.
9. The compound of claim 8 , wherein the QSY quencher is a hydrophilic QSY quencher.
10. The compound of claim 9 , wherein the hydrophilic QSY quencher is a sulfo-QSY quencher.
11. The compound of claim 8 , wherein the QC-1 quencher has the structure:
12. The compound of claim 1 having the formula (III):
wherein R comprises a QSY quencher or a QC-1 quencher; and
m and n are independently integers from 1 to 8.
13. The compound of claim 12 , wherein R is
and D is
14. A compound having a structure according to the following formula, or a pharmaceutically acceptable salt thereof:
15. The compound according to claim 14 , wherein the pharmaceutically acceptable salt has a structure according to the following formula:
16. A composition for use in labeling a protease in an animal comprising a compound of claim 1 and a pharmaceutically acceptable carrier.
17. A method of labeling a protease in an animal comprising the step of: administering the composition of claim 16 to the animal.
18. A method of visualizing a tumor in an animal comprising the steps of:
administering the composition of claim 16 to the animal; and
measuring a detectable signal generated in the animal from a reaction of the composition with a cathepsin cysteine protease;
where in the detectable signal is associated with a tumor in the animal.
19. The method of claim 18 , wherein the detectable signal is a fluorescent signal.
20. The method of claim 19 , wherein the fluorescent signal is generated at a tumor margin.