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, monophosphate, diphosphate, triphosphate, phosphoramidite ((R 2 N)(R′O)P) wherein R is linear, branched or cyclic alkyl and R′ is a protecting group, or H-phosphonate (HP(O)O—HNR 3 ), wherein R is linear, branched or cyclic alkyl;
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 an amido alkyl group;
Q is a detectable moiety; and,
M is a connecting group, and wherein n is an integer ranging from 0 to about 3.
2. The nucleic acid labeling compound of claim 1 , wherein A is H or H 4 O 9 P 1 − with an appropriate counterion.
3. The nucleic acid labeling compound of claim 1 , wherein X is O.
4. The nucleic acid labeling compound of claim 1 , wherein Y is OR 9 , wherein R 9 is H, alkyl or aryl.
5. The nucleic acid labeling compound of claim 1 , wherein Z is OR 10 , wherein R 10 is H, alkyl or aryl.
6. The nucleic acid labeling compound of claim 1 , wherein L is —C(O)NH(CH 2 ) n NH—, wherein n is an integer ranging from about 2 to about 10.
7. The nucleic acid labeling compound of claim 1 , wherein M is —NH(CH 2 ) n NH— or —C(O)(CH 2 ) n NH— wherein n is and integer from about 0 to about 5.
8. The nucleic acid labeling compound of claim 1 , wherein Q is biotin.
9. The nucleic acid labeling compound of claim 1 , wherein
A is H 4 O 9 P 3 − with an appropriate counterion;
X is O;
Y is H or OH;
Z is H or OH;
L is —C(O)NH(CH 2 ) 2 NH—,
M is —C(O)(CH 2 ) 5 NH—; and
Q is biotin.
10. The nucleic acid labeling compound of claim 1 , wherein the compound is a stereoisomer.
11. The nucleic acid labeling compound of claim 10 , wherein the compound has the following structure:
12. The nucleic acid labeling compound of claim 9 , wherein the compound has the following structure:
13. The nucleic acid labeling compound of claim 11 , wherein A is H or H 4 O 9 P 3 − with an appropriate counterion.
14. The nucleic acid labeling compound of claim 11 , wherein X is O.
15. The nucleic acid labeling compound of claim 11 , wherein Y is OR 9 , wherein R 9 is H, alkyl or aryl.
16. The nucleic acid labeling compound of claim 11 , wherein Z is OR 10 , wherein R 10 is H, alkyl or aryl.
17. The nucleic acid labeling compound of claim 11 , wherein L is —C(O) n H(CH 2 ) n NH—, wherein n is an integer ranging from about 2 to about 10.
18. The nucleic acid labeling compound of claim 11 , wherein M is —NH(CH 2 ) n NH— or —C(O)(CH2) n NH— wherein n is and integer from about 0 to about 5.
19. The nucleic acid labeling compound of claim 11 , wherein Q is biotin.
20. The nucleic acid labeling compound of claim 12 , wherein A is H or H 4 O 9 P 3 − with an appropriate counterion.
21. The nucleic acid labeling compound of claim 12 , wherein X is O.
22. The nucleic acid labeling compound of claim 12 , wherein Y is OR 9 , wherein R 9 is H, alkyl or aryl.
23. The nucleic acid labeling compound of claim 12 , wherein Z is OR 10 , wherein R 10 is H, alkyl or aryl.
24. The nucleic acid labeling compound of claim 12 , wherein L is —C(O)NH(CH 2 ) n NH—, wherein n is an integer ranging from about 2 to about 10.
25. The nucleic acid labeling compound of claim 12 , wherein M is —NH(CH 2 ) n NH— or —C(O)(CH 2 ) n NH— wherein n is and integer from about 0 to about 5.
26. The nucleic acid labeling compound of claim 12 , wherein Q is biotin.
27. A nucleic acid derivative produced by coupling a nucleic acid labeling compound according to claim 11 with a nucleic acid.
28. The nucleic acid derivative of claim 27 , wherein the coupling comprises incubating with a terminal transferase.
29. A hybridization product comprising the nucleic acid derivative according to claim 27 bound to a complementary probe.
30. The hybridization product of claim 29 , wherein the probe is attached to a solid support.
31. The hybridization product of claim 30 , wherein the solid support is glass.
32. A nucleic acid derivative produced by coupling a nucleic acid labeling compound according to claim 12 with a nucleic acid.
33. The nucleic acid derivative of claim 32 , wherein the coupling comprises incubating with a terminal transferase.
34. A hybridization product comprising the nucleic acid derivative according to claim 32 bound to a complementary probe.
35. The hybridization product of claim 34 , wherein the probe is attached to a solid support.
36. The hybridization product of claim 35 , wherein the solid support is glass.
37. A method of synthesizing a nucleic acid derivative comprising coupling a nucleic acid labeling compound according to claim 11 to a nucleic acid.
38. The method of claim 37 , wherein the coupling is achieved using a terminal transferase.
39. A method of detecting a nucleic acid comprising hybridizing a nucleic acid derivative according to claim 37 with a complementary probe.
40. The method according to claim 39 , wherein the probe is attached to a solid support.
41. The method according to claim 40 , wherein said solid support is glass.
42. A method of synthesizing a nucleic acid derivative comprising coupling a nucleic acid labeling compound according to claim 12 to a nucleic acid.
43. The method of claim 42 , wherein the coupling is achieved using a terminal transferase.
44. A method of detecting a nucleic acid comprising hybridizing a nucleic acid derivative according to claim 42 with a complementary probe.
45. The method according to claim 44 , wherein the probe is attached to a solid support.
46. The method according to claim 45 , wherein said solid support is glass.