Non-aggregating heptamethine cyanine fluorophores for in vivo imaging
Heptamethine cyanine fluorophore conjugates and conjugate precursors are disclosed. Methods of using the conjugates and conjugate precursors are also disclosed. The disclosed conjugates are neutral zwitterionic molecules and exhibit little or no aggregation.
1. A conjugate or conjugate precursor, or a stereoisomer thereof, according to Formula IA:
wherein p is 2, 3, or 4;
R a is —(R 1 ) q C(O)R c where q is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
each R b independently is C 1 -C 5 aliphatic;
R c is a targeting agent-containing group,
where y is an integer ≥1, or —OH;
R 1 is —CR 2 2 — where each R 2 independently is H, halo, optionally substituted aliphatic, or optionally substituted aryl;
R 3 and R 4 independently are aliphatic; and
R 13 to R 16 independently are aliphatic.
2. The conjugate or conjugate precursor according to claim 1 , wherein:
each R b independently is C 1 -C 5 alkyl;
R 1 is —CR 2 2 — where each R 2 independently is H, halo, optionally substituted alkyl, or optionally substituted aryl;
R 3 and R 4 independently are alkyl;
and
R 13 to R 16 independently are alkyl.
3. The conjugate or conjugate precursor according to claim 1 wherein R 1 is —CH 2 —.
4. The conjugate or conjugate precursor according to claim 1 , wherein:
each R b is the same;
R 3 and R 4 are the same; and
R 13 -R 16 are the same.
5. The conjugate or conjugate precursor according to claim 1 , wherein q is 2, 3, or 4.
6. The conjugate or conjugate precursor of claim 1 , wherein:
(i) R 3 and R 4 are methyl; or
(ii) R 13 -R 16 are methyl; or
(iii) both (i) and (ii).
7. The conjugate or conjugate precursor of claim 1 , wherein each R b is methyl.
8. The conjugate or conjugate precursor of claim 1 , wherein R a is —(CH 2 ) 3 C(O)R c where R c is a targeting agent-containing group and the conjugate is:
9. The conjugate of claim 1 , wherein R c is —N(H)Ab where Ab is an antibody.
10. The conjugate precursor of claim 1 , wherein R c is
and the conjugate precursor is:
11. A conjugate according to Formula IIA:
wherein p is 2, 3, or 4;
q is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
x is 2, 3, 4, 5, 6, 7, or 8;
R 1 is —CR 2 2 — where each R 2 independently is H, halo, optionally substituted aliphatic, or optionally substituted aryl;
R 3 and R 4 independently are C 1 -C 5 aliphatic;
R 13 to R 16 independently are C 1 -C 5 aliphatic;
each R b independently is C 1 -C 5 aliphatic; and
R c is a targeting agent-containing group.
12. The conjugate of claim 11 , wherein the conjugate is:
wherein x is 2, 3, 4, 5, 6, 7, or 8; and
R c is —N(H)Ab where Ab is an antibody.
13. A pharmaceutical composition, comprising:
a conjugate according to claim 1 ; and
a pharmaceutically acceptable carrier.
14. A method of imaging a subject or sample, comprising:
contacting a biological sample including or suspected of including a target with a conjugate according to claim 1 , wherein R c is a targeting agent-containing group, the targeting agent recognizing and binding to the target;
subsequently irradiating the biological sample with a quantity of light having a selected wavelength and selected intensity to induce fluorescence of the conjugate; and
detecting fluorescence of the irradiated biological sample, wherein fluorescence indicates presence of the target in the biological sample.
15. The method of claim 14 , wherein detecting fluorescence of the biological sample is performed ex vivo.
16. The method of claim 14 , wherein contacting the biological sample with the conjugate is performed in vivo by administering the conjugate or a pharmaceutical composition comprising the conjugate to a subject.
17. The method of claim 16 , wherein:
irradiating the biological sample comprises irradiating a target area of the subject; and
detecting fluorescence comprises obtaining an image of the irradiated target area, wherein fluorescence in the image indicates presence of the target in the target area.
18. The method of claim 17 , wherein the target is a tumor and the target area is an area in which the tumor is located.
19. The method of claim 18 , wherein the targeting agent recognizes and binds to cells of the tumor, irradiating the biological sample comprises irradiating the target area of the subject, and detecting fluorescence indicates presence of tumor cells in the target area, the method further comprising excising fluorescent tumor cells from the target area.
20. The method of claim 14 , wherein the targeting agent is an antibody.