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 .- 40 . (canceled)
41 . A compound of formula (II):
or a tautomer or a salt thereof, wherein:
Y is —OR y1 ;
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;
R B3 , R B4 , R C3 , and R C4 , at each occurrence, are independently H, C 1-10 alkyl, C 1-10 haloalkyl, C 3-12 cycloalkyl, —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; and
R y1 is H.
42 . The compound of claim 41 , or a tautomer or a salt thereof, wherein p is 1, 2, 3, or 4.
43 . The compound of claim 41 , or a tautomer or a salt thereof, wherein q is 1, 2, 3, 4, or 5.
44 . The compound of claim 41 , or a tautomer or a salt thereof, wherein q is 0.
45 . The compound of claim 41 , or a tautomer or a salt thereof, wherein p+q is 1 or 2.
46 . The compound of claim 41 , or a tautomer or a salt thereof, wherein p is 1 and R 3 is halogen.
47 . The compound of claim 41 , or a tautomer or salt thereof, wherein R 3 is fluorine.
48 . The compound of claim 41 , or a tautomer or salt thereof, wherein R 4 , at each occurrence, is halogen.
49 . The compound of claim 48 , or a tautomer or a salt thereof, wherein the halogen is fluorine.
50 . The compound of claim 41 , or a tautomer or salt thereof, wherein q is 1 or 2 and R 4 is fluorine.
51 . The compound of claim 41 , or a tautomer or a salt thereof, wherein q is 1 and R 4 is C 1-10 alkyl.
52 . The compound of claim 41 , wherein:
p+q is 1, 2, or 3;
R 3 , at each occurrence, is independently halogen or methyl; and
R 4 , at each occurrence, is independently halogen or methyl.
53 . The compound of claim 41 , or a tautomer or salt thereof, selected from the group consisting of:
54 . A kit comprising the compound of claim 41 , or a tautomer or a salt thereof.
55 . The kit of claim 54 , further comprising a luciferase.
56 . The kit of claim 54 , further comprising a buffer reagent.
57 . A method for detecting luminescence in a sample, the method comprising
contacting a sample with the compound of claim 41 , 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.
58 . The method of claim 57 , wherein the sample contains live cells.
59 . The method of claim 57 , wherein the sample contains a coelenterazine-utilizing luciferase.
60 . A method for detecting luminescence in a transgenic animal comprising
administering the compound of claim 41 , or a tautomer or a salt thereof, to a transgenic animal; and
detecting luminescence;
wherein the transgenic animal expresses a coelenterazine-utilizing luciferase.