Tumor-targeted optical contrast agents

• US 20040141920A1
• Filed: 09/03/2003
• Published: 07/22/2004
• Est. Priority Date: 01/18/2000
• Status: Active Grant

• Current Assignee: Mallinckrodt Incorporated (Mallinckrodt plc)
• Original Assignee: Mallinckrodt Incorporated (Mallinckrodt plc)
• Inventors: Rajagopalan, Raghavan, Dorshow, Richard B., Bugaj, Joseph E., Achilefu, Samuel I., Periasamy, Muthunadar P.
• Granted Patent: US 7,198,778 B2
• Time in Patent Office: Days
• US Class Current: 424/9.6
• CPC Class Codes: A61K 41/0033 Sonodynamic cancer therapy with sonochemically active agents or sonosensitizers, having their cytotoxic effects enhanced through application of ultrasounds ultrasound therapy per se A61N7/00
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: A61K 49/0056 Peptides, proteins, polyamino acids
• CPC Class Codes: C07D 209/14 Radicals substituted by nitrogen atoms, not forming part of a nitro radical
• CPC Class Codes: C07D 405/06 linked by a carbon chain containing only aliphatic carbon atoms
• CPC Class Codes: C07K 1/13 Labelling of peptides
• CPC Class Codes: C07K 14/655 Somatostatins

Abstract

Cyanine dye bioconjugates useful for diagnostic imaging and therapy are disclosed. The conjugates include several cyanine dyes with a variety of bis- and tetrakis (carboxylic acid) homologs. The compounds may be conjugated to bioactive peptides, carbohydrates, hormones, drugs, or other bioactive agents. The small size of the compounds allows more favorable delivery to tumor cells as compared to larger molecular weight imaging agents. The various dyes are useful over the range of 350 to 1300 nm, the exact range being dependent upon the particular dye. The use of dimethylsulfoxide helps to maintain the fluorescence of the compounds. The inventive compounds are useful for diagnostic imaging and therapy, in endoscopic applications for the detection of tumors and other abnormalities, for localized therapy, for photoacoustic tumor imaging, detection and therapy, and for sonofluoresence tumor imaging, detection and therapy.

Claims

1. A compound of formula

2. The compound of claim 1 wherein W⁴ and X⁴ are independently selected from the group consisting of —

NR³ and —

S—

;

Y⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

Z⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Dm, —

(CH₂)_a—

NHCO-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Dm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

A₂ is a single or a double bond;

B₂, C₂, and D₂ are independently selected from the group consisting of —

O—

, —

S—

, NR³, (CH2)_a—

CR¹R², and —

CR¹;

A₂, B₂, C₂, and D₂ may together form a 6- to 10-membered carbocyclic ring or a 6- to 10-membered heterocyclic ring optionally containing one or more oxygen, nitrogen, or sulfur atom;

a₄ and b₄ are independently from 0 to 3;

R¹ to R⁴, and R⁴⁵ to R⁵⁷ are independently selected from the group consisting of hydrogen, C₁-C₁₀ alkyl, C₅-C₁₂ aryl, C₁-C₁₀ alkoxyl, C₁-C₁₀ polyhydroxyalkyl, C₅-C₁₂ polyhydroxyaryl, C₁-C₁₀ aminoalkyl, mono- or oligosaccharide, peptide with 2 to 30 amino acid units, —

CH₂(CH₂OCH₂)_b—

CH₂—

OH, —

(CH₂)_a—

CO₂H, —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

OH and —

CH₂—

(CH₂OCH₂)_b—

CO₂H;

Bm and Dm are independently selected from the group consisting of a bioactive peptide containing 2 to 30 amino acid units, an antibody, a mono- or oligosaccharide, a glycopeptide, a metal chelating agent, a radioactive or nonradioactive metal complex, and an echogenic agent;

a and c are independently from 1 to 10; and

b and d are independently from 1 to 30.

3. The compound of claim 1 wherein each W⁴ and X⁴ is C(CH₃)₂;

Y⁴ is —

(CH₂)₂—

CONH-Bm;

Z⁴ is —

(CH₂)₂—

CONH-Dm;

A₂ is a single bond;

A₂, B₂, C₂, and D₂ together form a 6-membered carbocyclic ring;

each a₄ and b₄ is 1;

R⁴⁵ is galactose;

each R⁴⁶ to R⁵⁷ is hydrogen;

Bm is Octreotate;

Dm is bombesin (7-14).

4. The compound of claim 1 further comprising one to ten groups containing Bm, Dm, and combinations thereof wherein W⁴ and X⁴ additionally include CR¹R².

5. The compound of claim 4 wherein a porphyrin or a photodynamic therapy agent is attached to Bm, Dm, and combinations thereof.

38. A method of imaging a patient comprising administering a non-optical contrast agent composition further comprising the compound of claim 1 and performing at least one of an optical imaging procedure or a non-optical imaging procedure.

39. The method of claim 38 wherein the non-optical contrast agent composition is chosen from a magnetic resonance composition, a computed tomography composition, an x-ray composition, a nuclear imaging compostion, a positron emission tomography composition, a single photon emission computed tomography composition, and an ultrasound composition.

40. The method of claim 38 wherein the compound stabilizes or buffers the non-optical contrast agent composition.

6. A composition comprising a cyanine dye bioconjugate of formula

7. The composition of claim 6 wherein W⁴ and X⁴ are independently selected from the group consisting of —

NR³ and —

S—

;

Y⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

Z⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Dm, —

(CH₂)_a—

NHCO-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Dm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

A₂ is a single or a double bond;

B₂, C₂, and D₂ are independently selected from the group consisting of —

O—

, —

S—

, NR³, (CH2)_a—

CR¹R², and —

CR¹;

A₂, B₂, C₂, and D₂ may together form a 6- to 10-membered carbocyclic ring or a 6- to 10-membered heterocyclic ring optionally containing one or more oxygen, nitrogen, or sulfur atom;

a₄ and b₄ are independently from 0 to 3;

R¹ to R⁴, and R⁴⁵ to R⁵⁷ are independently selected from the group consisting of hydrogen, C₁-C₁₀ alkyl, C₅-C₁₂ aryl, C₁-C₁₀ alkoxyl, C₁-C₁₀ polyhydroxyalkyl, C₅-C₁₂ polyhydroxyaryl, C₁-C₁₀ aminoalkyl, mono- or oligosaccharide, peptide with 2 to 30 amino acid units, —

CH₂(CH₂OCH₂)_b—

CH₂—

OH, —

(CH₂)_a—

CO₂H, —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

OH and —

CH₂—

(CH₂OCH₂)_b—

CO₂H;

Bm and Dm are independently selected from the group consisting of a bioactive peptide containing 2 to 30 amino acid units, an antibody, a mono- or oligosaccharide, a glycopeptide, a metal chelating agent, a radioactive or nonradioactive metal complex, and an echogenic agent;

a and c are independently from 1 to 10; and

b and d are independently from 1 to 30.

8. The composition of claim 6 wherein each of W⁴ and X⁴ is C(CH₃)₂;

Y⁴ is —

(CH₂)₂—

CONH-Bm;

Z⁴ is —

(CH₂)₂-CONH-Dm;

A₂ is a single bond;

A₂, B₂, C₂, and D₂ together form a 6-membered carbocyclic ring;

a₄ and b₄ are independently galactose;

each of R⁴⁶ to R⁵⁷ is hydrogen;

Bm is Octreotate; and

Dm is bombesin (7-14).

9. The composition of claim 6 further comprising one to ten groups containing Bm, Dm, and combinations thereof wherein W⁴ and X⁴ additionally include CR¹R².

10. The composition of claim 6 wherein Bm, Dm, and combinations thereof further comprise a porphyrin or a photodynamic therapy agent.

11. The composition of claim 6 further comprising a non-optical contrast agent.

12. The composition of claim 6 wherein the agent is formulated as at least one of a liposome, a micelle, a microcapsule, or a microparticle.

13. The composition of claim 6 wherein the agent is formulated in a long-lived liposome.

14. A composition comprising indocyanine dye of Formula

15. A method of performing a diagnostic or therapeutic procedure comprising administering to an individual an effective amount of the composition of claim 14.

16. A method for performing a diagnostic or therapeutic procedure comprising

administering to an individual an effective amount of the compound of formula

wherein W⁴ and X⁴ are independently selected from the group consisting of —

O—

, —

NR³, and —

S—

;

Y⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

N(R³)—

(CH₂)_b—

CONH-Bm, (CH₂)_a—

N(R³)—

(CH₂)_c—

NHCO-Bm, —

(CH₂)_a—

N(R³)—

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

N(R³)—

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

CH₂—

(CH₂OCH₂)_d—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

CH₂—

(CH₂OCH₂)_d—

NHCO-Bm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

Z⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Dm, —

(CH₂)_a—

NHCO-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Dm, —

(CH₂)_a—

N(R³)—

(CH₂)_b—

CONH-Dm, (CH₂)_a—

N(R³)—

(CH₂)_c—

NHCO-Dm, —

(CH₂)_a—

N(R³)—

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Dm, —

(CH₂)_a—

N(R³)—

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

(CH₂)_a—

CONH-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

(CH₂)_a—

NHCO-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

CH₂—

(CH₂OCH₂)_d—

CONH-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

N(R³)—

CH₂—

(CH₂OCH₂)_d—

NHCO-Dm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

A₂ is a single or a double bond;

B₂, C₂, and D₂ are independently selected from the group consisting of —

O—

, —

S—

, —

Se—

, —

P—

, —

CR¹R², —

CR¹, alkyl, NR³, and —

C═

O;

A₂, B₂, C₂, and D₂ may together form a 6- to 12-membered carbocyclic ring or a 6- to 12-membered heterocyclic ring optionally containing one or more oxygen, nitrogen, or sulfur atom;

a₄ and b₄ are independently from 0 to 5;

R¹ to R⁴, and R⁴⁵ to R⁵⁷ are independently selected from the group consisting of hydrogen, C₁-C₁₀ alkyl, C₅-C₂₀ aryl, C1-C₁₀ alkoxyl, C₁-C₁₀ polyalkoxyalkyl, C₁-C₂₀ polyhydroxyalkyl, C₅-C₂₀ polyhydroxyaryl, C₁-C₁₀ aminoalkyl, glucose derivatives of R groups, cyano, nitro, halogen, saccharide, peptide, —

CH₂(CH₂OCH₂)_b—

CH₂—

OH, —

(CH₂)_a—

CO₂H, —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

OH and —

CH₂—

(CH₂OCH₂)_b—

CO₂H;

Bm and Dm are independently selected from the group consisting of a bioactive peptide, a protein, a cell, an antibody, an antibody fragment, a saccharide, a glycopeptide, a peptidomimetic, a drug, a drug mimic, a hormone, a metal chelating agent, a radioactive or nonradioactive metal complex, and an echogenic agent;

a and c are independently from 1 to 20; and

b and d are independently from 1 to 100, and performing the diagnostic or therapeutic procedure.

17. The method of claim 16 comprising administering to an individual an effective amount of the compound wherein W⁴ and X⁴ are independently selected from the group consisting of —

NR³ and —

S—

;

Y⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

Z⁴ is selected from the group consisting of —

(CH₂)_a—

CONH-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Dm, —

(CH₂)_a—

NHCO-Dm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Dm, —

(CH₂)_a—

NR³R⁴, and —

CH₂(CH₂OCH₂)_b—

CH₂NR³R⁴;

A₂ is a single or a double bond;

B₂, C₂, and D₂ are independently selected from the group consisting of —

O—

, —

S—

, NR³, (CH2)_a—

CR¹R², and —

CR¹;

A₂, B₂, C₂, and D₂ may together form a 6- to 10-membered carbocyclic ring or a 6- to 10-membered heterocyclic ring optionally containing one or more oxygen, nitrogen, or sulfur atom;

a₄ and b₄ are independently from 0 to 3;

R¹ to R⁴, and R⁴⁵ to R⁵⁷ are independently selected from the group consisting of hydrogen, C₁-C₁₀ alkyl, C₅-C₁₂ aryl, C₁-C₁₀ alkoxyl, C₁-C₁₀ polyhydroxyalkyl, C₅-C₁₂ polyhydroxyaryl, C₁-C₁₀ aminoalkyl, mono- or oligosaccharide, peptide with 2 to 30 amino acid units, —

CH₂(CH₂OCH₂)_b—

CH₂—

OH, —

(CH₂)_a—

CO₂H, —

(CH₂)_a—

CONH-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

CONH-Bm, —

(CH₂)_a—

NHCO-Bm, —

CH₂—

(CH₂OCH₂)_b—

CH₂—

NHCO-Bm, —

(CH₂)_a—

OH and —

CH₂—

(CH₂OCH₂)_b—

CO₂H;

Bm and Dm are independently selected from the group consisting of a bioactive peptide containing 2 to 30 amino acid units, an antibody, a mono- or oligosaccharide, a glycopeptide, a metal chelating agent, a radioactive or nonradioactive metal complex, and an echogenic agent;

a and c are independently from 1 to 10; and

b and d are independently from 1 to 30.

18. The method of claim 17 comprising administering to an individual an effective amount of the compound wherein each of W⁴ and X⁴ is C(CH₃)₂;

Y⁴ is —

(CH₂)₂—

CONH-Bm;

Z⁴ is —

(CH₂)₂—

CONH-Dm;

A₂ is a single bond;

A₂, B₂, C₂, and D₂ together form a 6-membered carbocyclic ring;

each a₄ and b₄ is 1;

R⁴⁵ is galactose;

each R⁴⁶ to R⁵⁷ is hydrogen;

Bm is Octreotate;

Dm is bombesin (7-14).

19. The method of claim 16 wherein said procedure utilizes light of wavelength in the region of 350-1300 nm.

20. The method of claim 16 wherein the diagnostic procedure is at least one of optical tomography and fluorescence endoscopy.

21. The method of claim 16 further comprising monitoring a blood clearance profile of said compound by a method selected from the group consisting of fluorescence, absorbance, and light scattering, wherein light of wavelength in the region of 350-1300 nm is used.

22. The method of claim 16 wherein said procedure further comprises imaging and therapy, wherein said imaging and therapy is selected from the group consisting of absorption, light scattering, photoacoustic and sonofluoresence technique.

23. The method of claim 16 wherein said procedure is capable of diagnosing atherosclerotic plaques and blood clots.

24. The method of claim 16 wherein said procedure comprises administering localized therapy.

25. The method of claim 16 wherein said therapeutic procedure comprises photodynamic therapy.

26. The method of claim 16 wherein said therapeutic procedure comprises laser assisted guided surgery for the detection of micrometastases.

27. The method of claim 16 further comprising adding a biocompatible organic solvent at a concentration of one to fifty percent of the compound to prevent in vivo or in vitro fluorescence quenching.

28. The method of claim 27 wherein said compound is dissolved in a medium comprising one to fifty percent dimethyl sulfoxide.

29. The method of claim 16 wherein the compound comprises one to ten groups containing Bm, Dm, and combinations thereof wherein W⁴ and X⁴ additionally include CR¹R² providing a cooperative effect to enhance binding of the compound.

30. The method of claim 29 further comprising attaching a compound selected from the group consisting of a porphyrin and a photodynamic therapy agent to Bm, Dm, and combinations thereof, and providing light of a wavelength sufficient to activate the porphyrin or phototherapy agent.

31. The method of claim 29 wherein the procedure monitors blood clearance of the compound to detect an abnormality.

32. The method of claim 29 further comprising activating the compound prior to performing the procedure.

33. The method of claim 16 further comprising administering a non-optical contrast agent and imaging by at least one of magnetic resonance, ultrasound, X-ray, positron emission tomography, computed tomography, and single photon emission computed tomography.

34. The method of claim 16 wherein the compound administered has at least one R group replaced by EDTA, DPTA, or DOTA.

35. The method of claim 34 wherein the compound administered further comprises a radioactive metal ion or a paramagnetic metal ion.

36. The method of claim 35 further comprising imaging by at least one of optical imaging and magnetic resonance imaging.

37. The method of claim 16 wherein the compound is administered in a formulation selected from at least one of liposomes, micelles, microcapsules, or microparticles.