Bioluminescent Endoscopy Methods And Compounds

• US 20090081129A1
• Filed: 08/29/2008
• Published: 03/26/2009
• Est. Priority Date: 08/29/2007
• Status: Active Grant

• Current Assignee: Biolume, Inc.
• Original Assignee: Biolume, Inc.
• Inventors: Bryan, Bruce, Neff, Philip T., Murphy, Randall B.
• Granted Patent: US 8,431,698 B2
• Time in Patent Office: Days
• US Class Current: 424/9.600
• CPC Class Codes: C07D 487/04 Ortho-condensed systems
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/763 Bioluminescence

Abstract

Bioluminescent endoscopy methods and compounds, wherein an anatomical object is examined by means of filling, perfusing, intubating, injecting, or otherwise administering a solution containing a bioluminescent substance or a mixture of luciferin and luciferase which produces bioluminescence, wherein a color or monochrome image of the object is constituted using the images and information based on bioluminescent emitted by the bioluminescent substance. Procedures are demonstrated which allow bioluminescent solutions to be perfused into certain tissue regions, such as but not limited to the common biliary duct, genitourinary tract, gastro-intestinal tract, cardiovascular system and lymphatic system wherein said structures may be conveniently visualized during surgery to avoid damage to these structures. Such images may also be combined with visual light images. Methods of detection of cancer cells using bioluminescence are provided. Preferred embodiments disclosed include membrane permeant coelenterazine analogs.

Claims

1. A membrane permeant coelenterazine analog having:

Structure II wherein R₄ and R₅ are independently alkyl, optionally substituted alkyl, aralkyl or optionally substituted aralkyl;

or

Structure III wherein p may be an integer from 1 to 20;

or

Structure IV wherein R₁ and R₂ are independently alkyl, optionally substituted alkyl, aryl, optionally substituted aryl, aralkyl, or optionally substituted aralkyl, and R₃ is independently alkyl or aralkyl;

or

Structure IV wherein r is an integer from 1 to 20.

2. The membrane permeant coelenterazine analog of claim 1 having Structure II, where R₅ is alkyl, optionally substituted alkyl, aralkyl, or optionally substituted alkakyl and R₄ is

—

(CH₂)_z—

O—

(CH₂)_y—

CH₃

wherein z and y are integers which are independently from 1 to 10.

3. The membrane permeant coelenterazine analog of claim 1 having Structure IV wherein:

R₁ and R₂ are independently

—

(CH₂)_z—

O—

(CH₂)_y—

CH₃

and wherein z and y are integers which are independently from 1 to 10 and R₃ is independently alkyl or aralkyl.

4. The membrane permeant coelenterazine analog of claim 1 having Structure IV wherein:

R₁ and R₂ are independently

—

(CH₂)_z—

O—

(CH₂)_y—

O—

(CH₂)_uCH₃

and wherein z, u and y are integers which are independently from 1 to 10, and R₃ is independently alkyl or aralkyl.

5. A compound of Structure II:

wherein R₄ and R₅ are independently alkyl, optionally substituted alkyl, aralkyl or optionally substituted aralkyl.

6. The compound of claim 5 where R₅ is alkyl, optionally substituted alkyl, aralkyl, or optionally substituted alkakyl and R₄ is

—

(CH₂)_z—

O—

(CH₂)_y—

CH₃

wherein z and y are integers which are independently from 1 to 10.

7. A compound of Structure III:

wherein p may be an integer from 1 to 20.

8. A compound of Structure IV:

wherein R₁ and R₂ are independently alkyl, optionally substituted alkyl, aryl, optionally substituted aryl, aralkyl, or optionally substituted aralkyl, and R₃ is independently alkyl or aralkyl.

9. The compound of claim 8 wherein R₁ and R₂ are independently

—

(CH₂)_z—

O—

(CH₂)_y—

CH₃

wherein z and y are integers which are independently from 1 to 10 and R₃ is independently alkyl or aralkyl.

10. The compound of claim 8 wherein R₁ and R₂ are independently

—

(CH₂)_z—

O—

(CH₂)_y—

O—

(CH₂)_u—

CH₃

wherein z, u and y are integers which are independently from 1 to 10 and R₃ is independently alkyl or aralkyl.

11. A compound of Structure V:

wherein r is an integer from 1 to 20.

12. A method of using a bioluminescence system to illuminate internal body structures of a subject.

13. The method of claim 12 wherein said method is used for illumination of one or more components of a circulatory system, nervous system, body organ, or lymphatic system of the subject.

14. The method of claim 13 wherein one component of the bioluminescence system is a membrane permeant coelenterazine analog.

16. The method of claim 14 wherein the cancerous cells are contained in a sentinel lymph node.

17. The method of claim 14 wherein one component of the bioluminescence system is a membrane permeant coelenterazine analog.

18. The method of claim 14 wherein one component of the bioluminescence system is combined with sugars or other molecules that are absorbed by cancer cells at a more rapid rate than the surrounding tissues.

15. A method of using a bioluminescence system to detect cancerous cells.

19. A method for detection of cancerous cells in a sentinel node biopsy, comprising:

injecting a luciferase into lymphatic tissues adjacent to a cancer tumor;

obtaining a biopsy sample of the lymphatic tissues;

treating the biopsy sample with coelenterazine or an analog of coelenterazine;

detecting bioluminescence using a camera, a luminometer, or by visual inspection.

20. A method of using a bioluminescence system post-operatively to illuminate a surgical tissue in order to inspect the surgical tissue.

21. A method of examining an animal, avian or human object, comprising the steps of:

perfusing, administering, injecting, superfusing, infiltrating, or otherwise applying a bioluminescent generating system to a selected animal, avian or human object to be examined to provide bioluminescent illumination to said object;

obtaining one or images of said object with bioluminescent illumination with a camera system sensitive to the wavelength of emission of the bioluminescent generating system.

22. The method of claim 21, wherein said one or more images are obtained in dim visible light conditions.

23. The method of claim 21, wherein said one or more images are color or monochrome images.

24. The method of claim 21, further comprising obtaining one or more images of said object with visible light illumination, said visible light comprising visible light in a wavelength range which does not include light in a wavelength range of light emitted by the bioluminescent generating system.

25. The method of claim 24, wherein said images are obtained with a camera system sensitive to the wavelength of emission of the visible light illumination.

26. The method of claim 25, wherein the visible light illumination is a red light illumination.

27. The method of claim 25, further comprising:

comparing said image with bioluminescent illumination with said image with visible light illumination.

28. The method of claim 27 further comprising a step of switching between said said image with bioluminescent illumination with said image with visible light illumination.

29. The method of claim 24, wherein the object comprises a component of a animal, avian or human circulatory system, nervous system, body organ, or lymphatic system.

30. The method of claim 29, wherein the object comprises gallbladder, ureter, lung, heart, or intestine.

31. The method of claim 21, wherein said images are obtained by an endoscopic examination of the object.

32. The method of claim 25, wherein said images are obtained by an endoscopic examination of the object.