Nucleic acid binding dyes and uses therefor
View Patent ↗The invention provides novel compounds and compositions of Formulas I and II, as well as methods of using them. The compounds can be used, for example, to quantify an amount of double stranded DNA in a sample subjected to nucleic acid amplification, or for real time monitoring of a nucleic acid amplification reaction. The compounds can be provided in a kit, for example, with other reagents and instructions for using the compounds and reagents.
1. A compound of Formula I:
wherein
R 1 , R 2 and R 3 are each H
Y is S or O;
W taken together with the atoms to which it is attached is a 6 membered heterocyclic ring;
n is 0;
Q is Q2:
wherein
Y is —CR 13 ═CR 14 —; m is 1 and p is 0;
R 6 is unsubstituted (C 1 -C 8 )alkyl, unsubstituted aryl, or unsubstituted (C 1 -C 8 )alkaryl;
R 9 , R 11 , and R 12 are each independently H, amino, or halo;
R 10 is H, methoxy, amino or halo;
one of R 13 and R 14 is H and the other is NR 15 R 16 ;
R 15 is (C 1 -C 6 )alkyl or (C 1 -C 6 )alkyl-NR 17 R 18 or C 3 H 7 N + (CH 3 ) 3 , wherein R 17 and R 18 are each independently H or (C 1 -C 6 )alkyl; and
R 16 is (C 1 -C 6 )alkyl-NR 17 R 18 or C 3 H 7 —N + (CH 3 ) 3 , wherein R 17 and R 18 are each independently H or (C 1 -C 6 )alkyl;
or a salt thereof.
2. The compound of claim 1 wherein R 1 , R 2 and R 3 are each H; Y is S; and n is 0.
3. The compound of claim 1 wherein Q is Q2; m is 1; R 6 is methyl, ethyl, propyl, phenyl, benzyl, 2-phenylethyl, or 3-phenylpropyl; R 9 , R 10 , R 11 , and R 12 are each H; R 13 is H; R 14 is NR 15 R 16 wherein R 15 is methyl, ethyl, propyl, or butyl.
4. The compound of claim 3 wherein R 16 is (C 1 -C 3 )alkyl-N(Me) 2 .
5. The compound of claim 1 wherein the compound is a compound of formula II:
wherein
R 6 is methyl, ethyl, propyl, phenyl, benzyl, 2-phenylethyl, or 3-phenylpropyl; and R 14 is
6. The compound of claim 1 wherein the compound displays enhanced emission in the presence of nucleic acids, in either in vitro or in vivo environments, compared to its emission in the same environment lacking nucleic acids.