Coelenterazine analogues
Described are coelenterazine analogues, methods for making the analogues, kits comprising the analogues, and methods of using the compounds for the detection of luminescence in luciferase-based assays.
1. A compound of formula (II)
or a tautomer or a salt thereof, wherein:
Y is nitro, or —NR y1 R y2 ;
p is 0, 1, 2, 3, or 4;
q is 0, 1, 2, 3, 4, or 5;
p+q is 1, 2, 3, 4, 5, 6, 7, 8, or 9;
R 3 , at each occurrence, is independently halogen, CN, nitro, C 1-10 alkyl, C 1-10 haloalkyl, —OR A3 , or —NR B3 R C3 ;
R 4 , at each occurrence, is independently halogen, CN, nitro, C 1-10 alkyl, C 1-10 haloalkyl, —OR A4 , or —NR B4 R C4 ;
R A3 and R A4 , at each occurrence, are independently H, C 1-10 haloalkyl, C 3-12 cycloalkyl, —C(O)C 1-10 alkyl, —C(O)C 3-12 cycloalkyl, or —C(O)C 1-3 alkylene-C 3-12 cycloalkyl, wherein each cycloalkyl is optionally substituted with 1-4 substituents independently selected from the group consisting of halogen, C 1-4 alkyl, and C 1-4 haloalkyl;
R B3 , R B4 , R C3 , and R C4 , at each occurrence, are independently H, C 1-10 haloalkyl, C 3-12 cycloalkyl, —C(O)C 1-10 alkyl, —C(O)C 3-12 cycloalkyl, —C(O)C 1-3 alkylene-C 3-12 cycloalkyl, —SO 2 C 1-10 alkyl, —SO 2 C 3-12 cycloalkyl, or —SO 2 C 1-3 alkylene-C 3-12 cycloalkyl, wherein each cycloalkyl is optionally substituted with 1-4 substituents independently selected from the group consisting of halogen, C 1-4 alkyl, and C 1-4 haloalkyl;
alternatively, R B3 and R C3 and/or R B4 and R C4 , together with the nitrogen atom to which each attaches form a 4- to 8-membered saturated or partially unsaturated heterocyclic ring, optionally substituted with 1-4 substituents independently selected from the group consisting of C 1-4 alkyl, C 1-4 haloalkyl, oxo, —OH, and —OC 1-4 alkyl;
R y1 and R y2 , at each occurrence, are independently H, C 1-10 alkyl, C 1-10 haloalkyl, C 3-12 cycloalkyl, or —C 1-3 alkylene-C 3-12 cycloalkyl, wherein each cycloalkyl is optionally substituted with 1-4 substituents independently selected from the group consisting of halogen, C 1-4 alkyl, and C 1-4 haloalkyl; and
alternatively, R y1 and R y2 together with the nitrogen atom to which each attaches form a 4- to 8-membered saturated or partially unsaturated heterocyclic ring, optionally substituted with 1-4 substituents independently selected from the group consisting of C 1-4 alkyl, C 1-4 haloalkyl, oxo, —OH, and —OC 1-4 alkyl.
2. The compound of claim 1 , or a tautomer or a salt thereof, wherein q is 0, 1, or 2.
3. The compound of claim 1 , or a tautomer or a salt thereof, wherein R y1 and R y2 are H.
4. The compound of claim 1 , or a tautomer or a salt thereof, wherein p+q is 1 or 2.
5. The compound of claim 1 , or a tautomer or a salt thereof, wherein p is 1 and R 3 is halogen.
6. The compound of claim 5 , or a tautomer or salt thereof, wherein R 3 is fluorine.
7. The compound of claim 1 , or a tautomer or salt thereof, wherein R 4 , at each occurrence, is halogen.
8. The compound of claim 7 , or a tautomer or a salt thereof, wherein the halogen is fluorine.
9. The compound of claim 1 , or a tautomer or salt thereof, wherein q is 1 or 2 and R 4 is fluorine.
10. The compound of claim 1 , or a tautomer or a salt thereof, wherein q is 1 and R 4 is C 1-10 alkyl.
11. The compound of claim 1 , or a tautomer or salt thereof, selected from the group consisting of:
12. A kit comprising the compound of claim 1 , or a tautomer or a salt thereof.
13. A method for detecting luminescence in a sample, the method comprising
contacting a sample with the compound of claim 1 , or a tautomer or a salt thereof;
contacting the sample with a coelenterazine-utilizing luciferase, if it is not present in the sample; and
detecting luminescence.
14. A method for detecting luminescence in a transgenic animal comprising
administering the compound of claim 1 , or a tautomer or a salt thereof, to a transgenic animal; and
detecting luminescence;
wherein the transgenic animal expresses a coelenterazine-utilizing luciferase.
15. The compound of claim 1 , or a tautomer or a salt thereof, wherein:
Y is nitro, or NR y1 R y2 ;
p is 0, 1, or 2;
q is 0, 1, or 2;
p+q is 1 or 2;
R 3 , at each occurrence, is independently halogen or C 1-4 alkyl; and
R 4 , at each occurrence, is independently halogen or C 1-4 alkyl.
16. The compound of claim 15 , or a tautomer or a salt thereof, wherein:
p is 0 or 1;
Y is NR y1 R y2 ;
R y1 and R y2 are H;
R 3 , at each occurrence, is independently fluorine or methyl; and
R 4 , at each occurrence, is independently fluorine or methyl.
17. A compound of formula:
or a tautomer or salt thereof.