Compounds and methods for assaying redox state of metabolically active cells and methods for measuring NAD(P)/NAD(P)H
View Patent ↗The present invention provides compounds and methods for assaying redox state of metabolically active cells and methods for assaying enzyme activity and/or metabolite level by coupling to redox defining co-factor NAD(P)/NAD(P)H measurement.
1. A compound is of Formula (IV):
wherein R 2 is selected from —CH 2 -aryl or —CH 2 -heteroaryl;
R 5 is selected from —CH 2 -aryl or —CH 2 -heteroaryl;
R 7 is selected from aryl or heteroaryl;
R 14 is H, C 1-4 alkyl, C 1-4 alkoxyl, C 2-4 hydroxylalkyl, C 2-4 alkoxyl, C 2-4 carboxylic acid, or C 2-4 amide;
R 9 and R 10 are independently selected from C 1-4 alkyl;
R 11 , R 12 and R 13 are independently selected from H, C 1-4 alkyl, C 1-4 alkoxyl, bromo, chloro or amino, or R 11 and R 12 can form a fused phenyl ring;
X is O, NH or a direct bond;
L is a direct bond or —C 6 (R 16 ) 4 CH 2 — or —(CH 2 ) m C(R 17 ) 2 (CH 2 ) n —Y—C(O)—;
R 16 is independently H, halogen, CH 3 , OCH 3 , or NO 2 ;
R 17 is independently H, C 1-4 alkyl; or both R 17 together can form an alkyl ring having from 3-7 carbons;
m is an integer from 0-2;
n is an integer from 0-2;
Y is O or NR 15 ; and
R 15 is H, C 1-4 alkyl, C 2-4 hydroxylalkyl, C 2-4 alkoxyl, C 2-4 carboxylic acid, or C 2-4 amide.
2. A kit comprising a compound according to claim 1 .
3. A compound is of Formula (II):
wherein R 1 is H, C 1-4 alkyl, C 1-4 hydroxylalkyl, C 3-7 cyclic ring, aryl, benzyl or substituted benzyl ring, heterocycle, heteroaryl and —(CH 2 ) n —P(Ph) 3 ;
R 4 and R 5 are independently selected from H, halogen, methyl, and trifluoromethyl;
R 14 is H, C 1-4 alkyl, C 1-4 alkoxyl, C 2-4 hydroxylalkyl, C 2-4 alkoxyl, C 2-4 carboxylic acid, or C 2-4 amide;
R 9 and R 10 are independently selected from C 1-4 alkyl;
R 11 , R 12 and R 13 are independently selected from H, C 1-4 alkyl, C 1-4 alkoxyl, bromo, chloro or amino, or R 11 and R 12 can form a fused phenyl ring;
X is O, NH or a direct bond;
L is a direct bond or —C 6 (R 16 ) 4 CH 2 — or —(CH 2 ) m C(R 17 ) 2 (CH 2 ) n —Y—C(O)—;
wherein at least one of X or L is not a direct bond;
R 16 is independently H, halogen, CH 3 , OCH 3 , or NO 2 ;
R 17 is independently H, C 1-4 alkyl or both R 17 together form an alkyl ring having from 3-7 carbons;
m is an integer from 0-2;
n is an integer from 0-2;
Y is O or NR 15 ; and
R 15 is H, C 1-4 alkyl, C 2-4 hydroxylalkyl, C 2-4 alkoxyl, C 2-4 carboxylic acid, or C 2-4 amide;
n′ is an integer from 2-7.
4. The compound according to Formula (III):
wherein R 4 and R 5 are independently selected from H, halogen, methyl, and trifluoromethyl;
R 14 is H, C 1-4 alkyl, C 1-4 alkoxyl, C 2-4 hydroxylalkyl, C 2-4 alkoxyl, C 2-4 carboxylic acid, or C 2-4 amide;
R 9 and R 10 are independently selected from C 1-4 alkyl;
R 11 , R 12 and R 13 are independently selected from H, C 1-4 alkyl, C 1-4 alkoxyl, bromo, chloro or amino, or R 11 and R 12 can form a fused phenyl ring;
X is O, NH or a direct bond;
L is a direct bond or —C 6 (R 16 ) 4 CH 2 — or —(CH 2 ) m C(R 17 ) 2 (CH 2 ) n —Y—C( 0 )—;
R 16 is independently H, halogen, CH 3 , OCH 3 , or NO 2 ;
R 17 is independently H, C 1-4 alkyl or both R 17 together form an alkyl ring having from 3-7 carbons;
m is an integer from 0-2;
n is an integer from 0-2;
Y is O or NR 15 ; and
R 15 is H, C 1-4 alkyl, C 2-4 hydroxylalkyl, C 2-4 alkoxyl, C 2-4 carboxylic acid, or C 2-4 amide.
5. A kit comprising a compound according to claim 3 .
6. A kit comprising a compound according to claim 4 .
7. A compound selected from the group consisting of: