Nucleic acid labeling compounds
View Patent ↗Nucleic acid labeling compounds containing heterocyclic derivatives are disclosed. The heterocyclic derivative containing compounds are synthesized by condensing a heterocyclic derivative with a cyclic group (e.g. a ribofuranose derivative). The labeling compounds are suitable for enzymatic attachment to a nucleic acid, either terminally or internally, to provide a mechanism of nucleic acid detection.
1. A nucleic acid labeling compound of the following structure:
wherein A is H or a functional group that permits the attachment of the nucleic acid labeling compound to a nucleic acid;
X is O, S, NR 1 or CHR 2 , wherein R 1 and R 2 are, independently, H, alkyl or aryl;
Y is H, N 3 , F, OR 9 , SR 9 or NHR 9 , wherein R 9 is H, alkyl or aryl;
Z is H, N 3 , F or OR 10 , wherein R 10 is H, alkyl or aryl;
L is amino alkyl;
Q is a detectable moiety; and
M is a connecting group, wherein m is an integer ranging from 0 to about 3.
2. A nucleic acid labeling compound according to claim 1 , wherein A is H or H 4 O 9 P 3 —; X is O; Y is H or OR 9 , wherein R 9 is H, alkyl or aryl; Z is H, N 3 , F or OR 10 , wherein R 10 is H, alkyl or aryl; L is —NH(CH 2 ) n NH—, wherein n is an integer ranging from about 2 to about 10; Q is biotin or carboxyfluorescein; and M is —CO(CH 2 ) 5 NH— or —CO(CH 2 ) 5 NHCO(CH 2 ) 5 NH—, wherein m is 1 or 0.
3. A nucleic acid labeling compound according to claim 2 , wherein Y is H or OH; Z is H or OH; L is —NH(CH 2 ) 4 NH—; Q is biotin; and m is 0.
4. A nucleic acid labeling compound according to claim 2 , wherein Y is H or OH; Z is H or OH; L is —NH(CH 2 ) 4 NH—; Q is 5-carboxyfluorescein; and m is 0.
5. A nucleic acid labeling compound according to claim 2 , wherein A is H; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is biotin; and m is 0.
6. A nucleic acid labeling compound according to claim 2 , wherein A is H 4 O 9 P 3 —; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is biotin; and m is 0.
7. A nucleic acid labeling compound according to claim 2 , wherein A is H; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is biotin; M is —CO(CH 2 ) 5 NH—; and m is 1.
8. A nucleic acid labeling compound according to claim 2 , wherein A is H 4 O 9 P 3 —; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is biotin; M is —CO(CH 2 ) 5 NH—; and m is 1.
9. A nucleic acid labeling compound according to claim 1 , wherein A is H; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is biotin; M is —CO(CH 2 ) 5 NH—; and m is 2.
10. A nucleic acid labeling compound according to claim 1 , wherein A is H 4 O 9 P 3 —; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is biotin; M is —CO(CH 2 ) 5 NH—; and m is 2.
11. A nucleic acid labeling compound according to claim 2 , wherein A is H; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is 5-carboxyfluorescein; and m is 0.
12. A nucleic acid labeling compound according to claim 2 , wherein A is H 4 O 9 P 3 —; Y is H; Z is H; L is —NH(CH 2 ) 4 NH—; Q is 5-carboxyfluorescein; and m is 0.
13. The nucleic acid labeling compound according to claim 6 , 8 , 10 or 12 having the formula:
wherein A, X, Y, Z, L, M, N, m and Q are as defined herein above.
14. The nucleic acid labeling compound according to claim 6 , 8 , 10 or 12 having the formula:
wherein A, X, Y, Z, L, M, N, m and Q are as defined herein above.
15. The nucleic acid labeling compound according to claim 6 , 8 , 10 or 12 having the formula:
wherein A, X, Y, Z, L, M, N, m and Q are as defined herein above.
16. A nucleic acid labeling compound of the following structure:
wherein A is H or a functional group that permits the attachment of the nucleic acid labeling compound to a nucleic acid;
X is O, S, NR 1 or CHR 2 , wherein R 1 and R 2 are, independently, H, alkyl or aryl;
Y is H, N 3 , F, OR 9 , SR 9 or NHR 9 , wherein R 9 is H, alkyl or aryl;
Z is H, N 3 , F or OR 10 , wherein R 10 is H, alkyl or aryl;
L is alkynyl alkyl;
Q is a detectable moiety; and
M is a connecting group, wherein m is an integer ranging from 0 to about 3.
17. A nucleic acid labeling compound according to claim 16 , wherein A is H or H 4 O 9 P 3 —; X is O; Y is H or OR 9 , wherein R 9 is H, alkyl or aryl; Z is H, N 3 , F or OR 10 , wherein R 10 is H, alkyl or aryl; L is —C≡CH 2 NH—; wherein n is an integer ranging from about 1 to about 10; Q is biotin or carboxyfluorescein; M is —CO(CH 2 ) 5 NH—; and m is 1 or 0.
18. A nucleic acid labeling compound according to claim 17 , wherein Y is H or OH; Z is H or OH; L is —C≡CH 2 NH—; Q is biotin; and m is 1.
19. A nucleic acid labeling compound according to claim 17 , wherein Y is H or OH; Z is H or OH; L is —C≡CH 2 NH—; Q is 5-carboxyfluorescein; and m is 1.
20. A nucleic acid labeling compound according to claim 17 , wherein A is H; Y is OH; Z is OH; L is —C≡CH 2 NH—; Q is biotin; M is —CO(CH 2 ) 5 NH—; and m is 1.
21. A nucleic acid labeling compound according to claim 17 , wherein A is H 4 O 9 P 3 —; Y is OH; Z is OH; L is —C≡CH 2 NH—; Q is biotin; M is —CO(CH 2 ) 5 NH—; and m is 1.
22. A nucleic acid labeling compound according to claim 17 , wherein A is H; Y is OH; Z is OH; L is —C≡CH 2 NH—; Q is 5-carboxyfluorescein; M is —CO(CH 2 ) 5 NH—; and m is 1.
23. A nucleic acid labeling compound according to claim 17 , wherein A is H 4 O 9 P 3 —; Y is OH; Z is OH; L is —C≡CH 2 NH—; Q is 5-carboxyfluorescein; M is —CO(CH 2 ) 5 NH—; and m is 1.
24. The nucleic acid labeling compound according to claim 21 or 23 having the formula:
wherein A, X, Y, Z, L, M, N, m and Q are as defined herein above.
25. The nucleic acid labeling compound according to claim 21 or 23 having the formula:
wherein A, X, Y, Z, L, M, N, m and Q are as defined herein above.
26. The nucleic acid labeling compound according to claim 21 or 23 having the formula:
wherein A, X, Y, Z, L, M, N, m and Q are as defined herein above.
27. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 2 to a nucleic acid.
28. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 13 to a nucleic acid.
29. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 14 to a nucleic acid.
30. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 15 to a nucleic acid.
31. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 17 to a nucleic acid.
32. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 24 to a nucleic acid.
33. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 25 to a nucleic acid.
34. A method of synthesizing a labeled nucleic acid comprising attaching a nucleic acid labeling compound according to claim 26 to a nucleic acid.