Dual-sensitizer-containing luminescent compounds, conjugates, and uses thereof
View Patent ↗The invention relates to novel luminescent compositions of matter containing two fluorophores (sensitizers), synthetic methods for making the compositions, macromolecular conjugates of the compositions, and the use of the compositions and their conjugates in various methods of detection. The invention also provides kits containing the compositions and their conjugates for use in the methods of detection.
1. A composition of matter comprising:
(i) a first sensitizer moiety having Formula (I) or (II);
(ii) a chelating moiety selected from EDTA, DTPA, TTHA, DOTA, TAGA, DOTP, DTPA-BMA, DO2P, and HP-DO3A covalently joined, optionally through a first linker independently selected from —NH— and a heterocyclic alkylene moiety having the formula
to the first sensitizer; and
(iii) a second sensitizer moiety having the Formula (II) covalently joined, through a second linker having the formula
to the chelating moiety of (ii);
wherein Formula (I) is:
and wherein:
X is CH—R 1 , S, or N—R 1 ;
R 1 is H; a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O;
R 2 is H; NH 2 ; carboxamide; hydrazide; acylhydrazide or alkylhydrazide; hydroxamate; COOH; CO—R′ or CO—O—R′, where R′ is a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O;
R 3 is a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, or alkylarene moieties are optionally further substituted with from 1-3 halo atoms; and
wherein the
at position 7 of the phenyl ring indicates the site of covalent attachment, optionally through the first linker, of the sensitizer moiety to the chelating moiety; and
wherein Formula (II) is:
and wherein:
Z is a CH—R 4 , S, or N—R 4 ;
R 4 is H; a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene, or alkylarene moieties are optionally further substituted with from 1-3 halo atoms;
R 5 is carboxamide; hydrazide; acylhydrazide or alkylhydrazide; hydroxamate; COOH; CO—R′ or CO—O—R′, where R′ is a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O; and
R 6 is H; a halogen; a linear alkylene (C 1 -C 20 ), a branched alkylene (C 3 -C 20 ), a cyclic alkylene (C 3 -C 10 ), a linear alkenylene (C 2 -C 20 ), a branched alkenylene (C 3 -C 20 ), a cyclic alkenylene (C 3 -C 10 ), a linear alkynylene (C 2 -C 20 ), a branched alkynylene (C 3 -C 20 ), a cyclic alkynylene (C 3 -C 10 ), an arene (C 6 -C 19 ), an alkylarene having a C 1 -C 20 alkyl portion and a C 6 -C 19 arene portion, a linear heteroalkylene (2-20 atoms), a branched heteroalkylene (3-20 atoms), or a cyclic heteroalkylene (3-10 atoms), wherein at least one atom of the heteroalkylene is N or O; and wherein the linear, branched, or cyclic alkylene, alkenylene, alkynylene, heteroalkylene, arene or alkylarene moieties are optionally further substituted with from 1-3 halo atoms; and wherein the
at position 7 of the phenyl ring indicates the site of covalent attachment, through the second linker, of the sensitizer moiety to the chelating moiety.
2. The composition of claim 1 , wherein X is N—R 1 .
3. The composition of claim 2 , wherein R 3 is CH 3 .
4. The composition of claim 3 , wherein said first sensitizer moiety has the formula:
5. The composition of claim 2 , wherein R 3 is CF 3 .
6. The composition of claim 5 wherein said first sensitizer moiety has the formula:
7. The composition of claim 2 , wherein R 1 is H.
8. The composition of claim 7 , wherein R 3 is CF 3 .
9. The composition of claim 8 , wherein said first sensitizer moiety has the formula:
10. The composition of claim 1 , wherein the Z is N—R 4 .
11. The composition of claim 10 , wherein R 4 is a cyclopropyl moiety.
12. The composition of claim 11 , wherein R 5 is an organic acid moiety having the formula COOH.
13. The composition of claim 11 , wherein R 6 is an F atom.
14. The composition of claim 13 , wherein said second sensitizer moiety has the formula:
15. The composition of claim 1 , wherein the chelating moiety is selected from the group consisting of: TTHA, DOTA, TAGA, DOTP, DTPA-BMA, DO2P, and HP-DO3A.
16. The composition of claim 1 , wherein the chelating moiety is EDTA.
17. The composition of claim 1 , wherein the chelating moiety is DTPA.
18. The composition of claim 1 , wherein one of the sensitizer moieties is covalently linked at the R 2 or R 5 position, optionally through a third linker moiety, to a conjugating group.
19. The composition of claim 18 , wherein the conjugating group is selected from S═C═N— and Br—CH 2 —CO—.
20. The composition of claim 1 further comprising a metal ion chelated to the chelating moiety.
21. The composition of claim 20 , wherein the metal is a lanthanide selected from the group consisting of: Gd(III), Dy(III), Ho(III), Er(III), Eu(III), Tb(III), Sm(III), Ce(III), Pr(III), Yb(III), Tm(III), Nd(III), and Tb(IV).
22. The composition of claim 21 , wherein the lanthanide is Tb(III).
23. The composition of claim 21 , wherein the lanthanide is Eu(III).
24. The composition of claim 21 , wherein the lanthanide is Dy(III).
25. The composition of claim 21 , wherein the lanthanide is Sm(III).
26. The composition of claim 1 , wherein the composition is conjugated to a macromolecule.
27. The composition of claim 26 , wherein the macromolecule is a polypeptide.
28. The composition of claim 27 , wherein the polypeptide is an antibody or antigen-binding fragment thereof.
29. The composition of claim 27 , wherein the polypeptide is a ligand for a cellular receptor.
30. The composition of claim 27 , wherein the macromolecule is a nucleic acid.
31. The composition of claim 30 , wherein the nucleic acid is RNA.
32. The composition of claim 31 , wherein the nucleic acid is DNA.
33. A pharmaceutical composition comprising the composition of claim 1 and a pharmaceutically acceptable carrier.
34. A pharmaceutical composition comprising the composition of claim 1 and a pharmaceutically acceptable carrier, wherein the composition of claim 1 is conjugated to an antibody or antigen-binding fragment thereof.
35. The composition of claim 13 , wherein both of the first and second sensitizer moieties have the formula: