Disinfection of biological fluids using asymmetric cyanine dyes

• US 20060257844A1
• Filed: 05/10/2006
• Published: 11/16/2006
• Est. Priority Date: 05/13/2005
• Status: Active Grant

• Current Assignee: American National Red Cross
• Original Assignee: American National Red Cross
• Inventors: Wagner, Stephen J.
• Granted Patent: US 7,678,538 B2
• Time in Patent Office: Days
• US Class Current: 435/2
• CPC Class Codes: A01N 43/78 1,3-Thiazoles; Hydrogenated 1,3-thiazoles
• CPC Class Codes: A01N 61/00 Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action

Abstract

Asymmetric cyanine dyes of Formula I bind nucleic acid but not red blood cell membrane, and function as photosensitizers when rigidly bound but not when free in solution. Unbound dye thus causes minimal oxidative damage. The dyes do not substantially accumulate in red blood cells, thereby minimizing hemolysis due to oxidative damage. Biological fluids can be disinfected by mixing the fluid with these asymmetric cyanine dye that binds to nucleic acid, irradiating the mixture, and recovering clinically significant components from the biological fluid and/or assaying the fluid for pathogens.

Claims

1. A method for disinfecting a biological fluid comprising:

(a) creating a mixture of the biological fluid and a nucleic acid binding, cell-membrane-permeable, asymmetric cyanine dye;

(b) irradiating the resulting mixture to achieve a viral inactivation capacity corresponding to at least about 7 Log₁₀ VSV; and

(c) assaying the biological fluid for a pathogen.

2. The method of claim 1, wherein the cyanine dye has the structure of Formula I:

3. The method of claim 2, wherein R¹ and R³ are saturated or unsaturated hydrocarbons of C₃ or fewer, n, m, and p are 0, and W is a fused bicyclic heteroaryl moiety.

4. The method of claim 2, wherein the asymmetric cyanine dye is thiazole orange.

5. The method of claim 1, further comprising the step of assaying the irradiated mixture for a virus.

6. The method of claim 1, wherein the biological fluid comprises red blood cells.

7. The method of claim 1, wherein the asymmetric cyanine dye in the mixture is at least about 50 μ

M.

8. The method of claim 1, further comprising the step of incubating the mixture prior to irradiation for about 5 to about 60 minutes.

9. The method of claim 2, wherein the cyanine dye-containing biological fluid is irradiated with white light.

10. The method of claim 2, wherein the thiazole orange-containing biological fluid is irradiated with about 5 to about 10 J/cm² white light.

11. The method of claim 6, wherein the method effects no more than about 1.0% hemolysis on storage at 42 days following irradiation.

12. The method of claim 6, wherein potassium efflux from the red blood cells is less than 30 mM on storage at four days following irradiation.

13. A method for disinfecting a biological fluid containing red blood cells comprising:

(a) creating a mixture of the biological fluid containing red blood cells and a nucleic acid binding, cell-membrane-permeable asymmetric cyanine dye;

(b) irradiating the resulting mixture; and

(c) recovering intact red blood cells from the mixture.

14. The method of claim 13, wherein the mixture is irradiated with at least about 5 to about 10 J/cm² white light.

15. The method of claim 13, wherein the method effects no more than about 1.0% hemolysis on storage at 42 days following irradiation.

16. The method of claim 13, wherein potassium efflux from the red blood cells is less than 30 mM on storage at four days following irradiation.

17. The method of claim 13, wherein the asymmetric cyanine dye in the mixture is at least about 50 μ

M.

18. The method of claim 13, further comprising the step of assaying the irradiated mixture for a virus.

19. The method of claim 13, wherein the asymmetric cyanine dye is added to the mixture at about 80 μ

M;

the mixture is incubated prior to irradiation for about 5 to about 60 minutes;

the mixture is irradiated with at least about 5 J/cm² white light; and

the resulting mixture is assayed for a pathogen.

20. The method of claim 13, wherein the cyanine dye has the structure of Formula I:

21. The method of claim 20, wherein R¹ and R³ are saturated or unsaturated hydrocarbons of C₃ or fewer;

n, m, and p are 0, and W is a fused bicyclic heteroaryl moiety.