Dual-sensitizer-containing luminescent compounds, conjugates, and uses thereof

1. A method of detecting a target with a probe, the method comprising:

contacting a sample with, or delivering to a subject, a luminescent probe composition comprising a targeting-probe moiety having an affinity for a target, the targeting-probe moiety covalently conjugated to a luminescent chelate composition comprising:

(i) a first sensitizer moiety;

(ii) a chelating moiety covalently joined, optionally through a first linker, to the first sensitizer; and

(iii) a second sensitizer moiety covalently joined, optionally through a second linker, to the chelating moiety of (ii);

wherein, the first and second sensitizer moieties, independently, have the Formula (I) or the Formula (II), wherein Formula (I) is:

and wherein:

X is CH—R 1 , O, 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 a linker, of the sensitizer moiety to the chelating moiety; and

wherein Formula (II) is:

and wherein:

Z is CH—R 4 , O, 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, 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, optionally through a linker, of the sensitizer moiety to the chelating moiety; and

wherein the chelating moiety is selected from the group consisting of: EDTA, DTPA, TTHA, DOTA, TAGA, DOTP, DTPA-BMA, DO2P, and HP-DO3A; and

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; and

wherein the luminescent composition further comprises a lanthanide chelated to the chelating moiety, the 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); and

detecting a signal produced from the luminescent probe composition.

2. The method of claim 1 , wherein the sample comprises one or more cells.

3. The method of claim 1 , wherein the first and the second linkers are independently —NH— or a heterocyclic alkylene.

4. The method of claim 1 , wherein the detection comprises detecting luminescence emissions from the luminescent probe composition.

5. The method of claim 1 , wherein the detection comprises detecting fluorescence emissions from one or both sensitizers of the luminescent probe composition.

6. The method of claim 1 , wherein X is N—R 1 .

7. The method of claim 6 , wherein R 3 is CH 3 .

8. The method of claim 7 , wherein the first or second sensitizer moiety has, or the first and second sensitizer moieties have, the formula:

9. The method of claim 6 , wherein R 3 is CF 3 .

10. The method of claim 9 , wherein the first or second sensitizer moiety has, or the first and second sensitizer moieties have, the formula:

11. The method of claim 1 , wherein R 1 is H.

12. The method of claim 11 , wherein R 3 is CF 3 .

13. The method of claim 12 , wherein the first or second sensitizer moiety has, or the first and second sensitizer moieties have, the formula:

14. The method of claim 1 , wherein the Z is N—R 4 .

15. The method of claim 14 , wherein R 4 is a cyclopropyl moiety.

16. The method of claim 15 , wherein R 5 is an organic acid moiety having the formula COOH.

17. The method of claim 15 , wherein R 6 is an F atom.

18. The method of claim 16 , wherein R 6 is an F atom.

19. The method of claim 17 , wherein the first or second sensitizer moiety has, or the first and second sensitizer moieties have, the formula:

20. The method of claim 1 , wherein the chelating moiety is selected from the group consisting of: EDTA, DTPA, TTHA, DOTA, TAGA, DOTP, DTPA-BMA, DO2P, and HP-DO3A.

21. The method 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.

22. The method of claim 21 , wherein the conjugating group is selected from S═C═N— and Br—CH 2 —CO—.

23. The method of claim 1 , wherein the luminescent chelate composition is conjugated to a macromolecule.

24. The method of claim 23 , wherein the macromolecule is a polypeptide.

25. The method of claim 24 , wherein the polypeptide is an antibody or antigen-binding fragment thereof.

26. The method of claim 24 , wherein the polypeptide is a ligand for a cellular receptor.

27. The method of claim 24 , wherein the macromolecule is a nucleic acid.