Class of stable heptamethine cyanine fluorophores and biomedical applications thereof

• US 10,280,307 B2
• Filed: 11/05/2014
• Issued: 05/07/2019
• Est. Priority Date: 11/05/2014
• Status: Active Grant

• Current Assignee: Department of Health and Human Services (State of Victoria)
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Schnermann, Martin John, Nani, Roger Rauhauser, Shaum, James Blaine
• Primary Examiner(s): Hartley, Michael G.
• Assistant Examiner(s): Samala, Jagadishwar R
• Application Number:
  US15/524,567
• Publication Number:
  US 20180273758A1
• Time in Patent Office: 1,644 Days
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: C07D 209/10 with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
• CPC Class Codes: C07D 209/12 Radicals substituted by oxygen atoms
• CPC Class Codes: C07D 209/14 Radicals substituted by nitrogen atoms, not forming part of a nitro radical
• CPC Class Codes: C09B 23/086 more than five >CH- groups
• CPC Class Codes: C09K 11/06 containing organic luminescent materials
• CPC Class Codes: C09K 2211/1018 Heterocyclic compounds
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

Embodiments of C4′-alkyl-ether heptamethine cyanine fluorophores according to general formula I, and pharmaceutically acceptable salts thereof, are disclosed. Methods of making and using the C4′-alkyl-ether heptamethine cyanine fluorophores also are disclosed.

Claims

1. A compound, or a pharmaceutically acceptable salt thereof, of formula I:

2. The compound of claim 1, wherein R³ is a biomolecule-containing group.

3. The compound of claim 2, wherein the biomolecule is an antibody, a peptide, a protein, an amino acid, a nucleoside, a nucleotide, a nucleic acid, an oligonucleotide, a carbohydrate, a lipid, a hapten, or a receptor ligand.

4. The compound of claim 1, wherein R³ is

5. The compound of claim 4, wherein R^b is an antibody, a peptide, a protein, an amino acid, a nucleic acid, nucleotide, an oligonucleotide, a lipid, a hapten, or a receptor ligand.

6. The compound of claim 1, wherein each R^a independently is hydrogen or halo, and m is 2 or 3.

7. The compound of claim 1, wherein R¹⁴ to R¹⁷ are methyl.

8. The compound of claim 1, wherein n is 2.

9. The compound of claim 1, wherein:

R⁵, R⁶, R⁸, R¹⁰, R¹¹ and R¹³ are hydrogen;

R⁴ and R⁹ independently are lower alkyl or a sulfonate-containing group; and

R⁷ and R¹² independently are hydrogen, a sulfonate-containing group, or a trialkyl amino group.

10. The compound of claim 9, wherein:

R⁴ and R⁹ are n-propyl or —

(CH₂)₄SO₃⁻

; and

R⁷ and R¹² are hydrogen or —

SO₃⁻

Na⁺.

11. The compound of claim 1, having the general formula:

12. The compound of claim 1, wherein:

n is 2;

R¹ is —

CH₂—

;

m is 2;

R² is methyl;

R¹⁴ to R¹⁷ are methyl;

R⁵, R⁶-R⁸ and R¹¹-R¹³ are hydrogen;

R⁴ and R⁹ are n-propyl;

Z is halide, and

R³ is

13. The compound of claim 1, wherein:

n is 2;

R¹ is —

CH₂—

;

m is 2;

R² is methyl;

R¹⁴ to R¹⁷ are methyl;

R⁵, R⁶, R⁸, R¹⁰, R¹¹ and R¹³ are hydrogen;

R⁴ and R⁹ are —

(CH₂)₄SO₃⁻

;

R⁷ and R¹² are —

SO₃⁻

;

Z is an alkali metal cation, and

R³ is

14. A method, comprising:

contacting a biological sample with a compound of claim 1;

irradiating the biological sample by application of light having a wavelength or range of wavelengths in the near-infrared range; and

detecting fluorescence of the irradiated biological sample, wherein fluorescence indicates presence of the compound in the biological sample.

15. The method of claim 14, wherein detecting fluorescence comprises obtaining a fluorescence-based image of the irradiated biological sample.

16. The method of claim 15, wherein the compound comprises a biomolecule capable of binding to a target suspected of being present within the biological sample and fluorescence indicates the target is present in the biological sample, the method further comprising:

removing unbound compound from the biological sample prior to obtaining the image.

17. The method of claim 14, wherein the biological sample comprises cells in solution and the compound comprises a moiety capable of binding to at least some cells in the solution, the method further comprising performing flow cytometry to separate cells to which the compound has bound from cells to which the compound did not bind.

18. The method of claim 14, wherein contacting the biological sample with the compound is performed in vivo by administering the compound to a subject.

19. The method of claim 18, wherein the compound comprises a biomolecule capable of binding to a target suspected of being present within the biological sample.

20. The method of claim 19, wherein the target is an antigen, and the compound comprises an antibody capable of recognizing and binding to the antigen.

21. The method of claim 19, wherein:

irradiating the biological sample comprises irradiating a target area of the subject with near-infrared radiation; 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.

22. The method of claim 19, wherein the target is a tumor and the target area is an area in which the tumor is located.

23. The method of claim 22, wherein the biomolecule is capable of recognizing and binding to cells of the tumor, irradiating the biological sample comprises irradiating the target area of the subject with near-infrared radiation, and detecting fluorescence indicates presence of tumor cells in the target area, the method further comprising:

excising fluorescent tumor cells from the target area.

24. The method of claim 18, wherein the compound comprises a drug, the biological sample is a bodily fluid or tissue, and detecting fluorescence of the irradiated biological sample indicates presence of the drug in the biological sample.

25. The method of claim 18, wherein detecting fluorescence of the biological sample is performed ex vivo.